KR20090075481A - Antioxidant composition comprising glycyrrhiza glabra extract - Google Patents

Abstract

An antioxidation composition containing a Glycyrrhiza glabra extract with excellent antioxidation activity is provided to improve stability without side effect and raise the contents of 7-O-methylluteone and dehydroglyasperin C. An antioxidation composition comprises a Glycyrrhiza glabra extract. The Glycyrrhiza glabra extract is obtained using C1-5 alcohol, hexane, dichloromethane, methylene chloride, chloroform, ethylacetate or ethylether. A solvent for extracting is mixture solvent of methylene chloride or normal hexane and ethanol in a ratio of 10:1-8:1. The Glycyrrhiza glabra extract comprises 0.4-0.6 weight part of 7-O-methyllutheone and 0.2-0.4 weight part of dehydroglyasperin C based on 100 weight part.

Description

Antioxidant composition comprising licorice extract {ANTIOXIDANT COMPOSITION COMPRISING GLYCYRRHIZA GLABRA EXTRACT}

The present invention relates to an antioxidant composition comprising a licorice extract excellent in antioxidant capacity as an active ingredient and a method for preparing the licorice extract having enhanced antioxidant capacity.

Most organisms are subjected to various environmental stresses caused by biological stresses such as germs, pests or viruses, or deterioration of the global environment. When a variety of stresses are applied to living organisms, oxygen, which is essential for life, is converted into reactive oxygen species (ROS), which are highly reactive and cause serious physiological disorders. The reactive oxygen species is recognized as a cause for promoting aging, and has been reported to cause a variety of diseases.

Specifically, the reactive oxygen species or free radicals destroy cells, cut off the connective tissue of the dermal layer of the skin, or cause cross-linking, thereby causing skin-related abnormalities such as wrinkles, atopic dermatitis, acne or skin cancer, as well as cancer and myocardial infarction. cardiovascular diseases, including myocardial infarction, stroke, and atherosclerosis, degenerative neurological diseases such as amyotrophic laternal sclerosis or Parkinson's disease (PD) (chronic neurodegenerative disease), rheumatoid arthritis and other causes have been reported.

In addition, reactive oxygen species can produce a harmful substance called lipid peroxide by peroxidating lipids in the human body, and the lipid peroxide acts on blood vessels and causes various adult diseases such as arteriosclerosis and blood serum.

It is a system that removes these active oxygen species in the living body, such as superoxide dismutase (SOD, EC 1. 15. 1.1), peroxidase (POD), catalase (CAT), etc. Polymeric antioxidant enzymes and antioxidants such as vitamin C, vitamin E or glutathione.

However, due to pollution, ultraviolet rays or various causes including the biological stress and aging, the antioxidant system of the human body gradually becomes unable to operate normally, and in this case, the active oxygen species can be removed to protect by ingesting antioxidants. There is a need for an in vivo supply of a substance that is capable of antioxidants.

Conventionally, synthetic antioxidants having excellent antioxidant power have been reported, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) or tertiary butylhydropuinone (TBHQ).

However, the synthetic antioxidants are toxic to some of the absorbent substances in the body, can be teratogenic factors and carcinogenous material (Ito, et . Al ., 1983), causing hepatomegaly or enzyme activity in the liver ( Safety issues such as increasing microsomal enzyme activity and causing pathological damage to organ tissues such as lungs and kidneys (Branen, 1975) have been raised.

Therefore, in recent years, many studies have been conducted to develop an antioxidant that is safer to the human body and excellent in antioxidant power. As part of this research, many studies on natural antioxidants are underway. More specifically, with the growing awareness of modern people's safety and health needs, natural antioxidants have been accumulating, as studies have reported that food intake, including natural antioxidants, can reduce the incidence of diseases caused by reactive oxygen species. There is a growing interest in.

As a result of continuous research on natural antioxidants, such as searching for antioxidant effects on natural plants, many natural antioxidants have been reported.

Natural antioxidants reported to date include tocophenols, flavono derivatives, sesamol, gossypol as natural antioxidants, isoflavones, phenolic acids, tocopherols, and phytic acid. ), Saponins and trypsin inhibitors have been reported as antioxidants (Hayes et . Al ., 1977).

Along with these findings, studies on natural extracts containing high amounts of antioxidants or excellent anti-oxidant properties, which can be used as foods or included in foods, for the prevention of diseases caused by reactive oxygen species and prevention of aging Or the demand for development is increasing.

In order to solve the problems of the prior art, an object of the present invention is to provide an antioxidant composition comprising a licorice extract excellent in antioxidant capacity as an active ingredient.

In addition, the present invention, 7- O In order to solve the problems of the prior art and an object thereof is to provide an antioxidant composition comprising a base Theon methyl (7- O -methylluteone) as an active ingredient.

In addition, the present invention, 7- O from licorice to solve the above problems - is an object of the present invention to provide a process for purifying methyl Lu Theon (7- O -methylluteone) or dihydro article ria peorin C (Dehydroglyasperin C) .

In order to achieve the above object, the present invention provides an antioxidant composition comprising a licorice extract excellent in antioxidant capacity as an active ingredient. The composition may preferably be a food composition. Examples of the food composition of the present invention include dietary supplements, food additives or beverages.

The present invention is 7- O excellent antioxidant ability to attain the object - to provide an antioxidant composition comprising a base Theon methyl (7- O -methylluteone) as an active ingredient.

In addition, the present invention provides a method for purifying 7- O -methylrutheon or dihydrogliasperin C from licorice to achieve the above object.

Hereinafter, the present invention will be described in more detail.

The inventors of the present invention, among the licorice with a variety of origin, the licorice of the country of origin, that is, when the domestic licorice is extracted using a mixed solvent mixed with methylene chloride or normal hexane and ethanol (9: 1), 1,1- in the extract diphenyl-2-picryl the hydroxyl using a Brazilian (1,1'-dephenyl-2-picrylhydrazyl , DPPH) found to be excellent in the antioxidant activity by the method of measuring the DPPH scavenging 7-O - methyl Lu Theon (7- O -methylluteone), Dehydroglyasperin C (Dehydroglyasperin C) and Dehydroglyasperin D (Dehydroglyasperin D) was confirmed that the content of the markedly increased to complete the present invention, based on this it was completed the present invention .

The present invention relates to an antioxidant composition comprising a licorice extract excellent in antioxidant capacity as an active ingredient.

The licorice is a perennial herb belonging to the Rosacea legume, and is a representative plant that uses different kinds of plants from among plants used as natural herbs. The licorice may be preferably Glycyrrhiza glabra . In the present invention, the Spanish acid licorice is licorice ( Glycyrrhiza) that is a related plant of the uralensis ) or Siberian licorice ( Glycyrrhiza) glabra var . glandifera ) was found to have superior antioxidant activity. In addition, the licorice may be any one or more selected from the group consisting of leaves, stems and roots, preferably may be one or more selected from the group consisting of roots and stems.

In addition, the licorice may be preferably domestic licorice. In the present invention, the domestic licorice refers to licorice, which is a production area of licorice, that is, a plantation or harvesting place of licorice. In the present invention, even if the plantation is different, the components of the licorice extract is different, especially the components related to the antioxidant activity was found to be remarkably different, especially compared to licorice produced domestically in China or Russia Remarkably excellent antioxidant activity, and the amount of 7- O- methylluteone, Dehydroglyasperin C and Dehydroglyasperin D, which were found to be excellent in antioxidant activity as a measure of DPPH scavenging activity in domestic licorice, was significantly higher than licorice grown in other countries Was confirmed to be high.

Licorice extract of the present invention can be prepared by a conventional plant extraction method using an extraction solvent.

The extraction solvent may be an alcohol having 1 to 5 carbon atoms, hexane, dichloromethane, methylene chloride, chloroform, ethyl acetate, ethyl ether, or a mixed solvent thereof, preferably a mixed solvent of methylene chloride or normal hexane and ethanol More preferably, a mixed solvent of normal hexane and ethanol. The content ratio of the mixed solvent of normal hexane and ethanol may be preferably 8: 1 to 10: 1 (normal hexane: ethanol) on a volume ratio basis. In order to obtain the main component showing the antioxidant activity in high yield, considering the properties of the material showing the antioxidant activity and the extraction yield of the extract, the content ratio of methylene chloride or a mixed solvent of normal hexane and ethanol is 8: 1 by volume ratio A mixed solvent of normal hexane and ethanol, which is from 10: 1 (normal hexane: ethanol), is preferable.

Licorice extract of the present invention may be prepared according to a conventional method for producing a plant extract, preferably may be a pressure extraction method or an ultrasonic grinding extraction method, more preferably a pressure extraction method.

The pressure extraction method may be a device extraction method using a device. The pressure extraction method may be carried out at 40 to 60 ℃ and 1400 psi to 1600 psi conditions, preferably at 45 to 55 ℃ and 1450 psi to 1550 psi conditions. In addition, the pressure extraction method may be performed for 5 minutes to 10 hours, preferably 10 minutes to 1 hour.

The ultrasonic grinding extraction method may be carried out at 40 to 60 ℃, preferably 45 to 55 ℃ and 5 to 100 kHZ, preferably 10 to 75 kHZ, more preferably 20 to 40 kHZ. In addition, the pressure extraction method may be performed for 1 day to 48 days, preferably 10 days to 30 days, more preferably 14 days to 21 days.

Extraction conditions and extraction time of the extraction method is a range determined in order to obtain an extract in high yield, to increase the content of the active ingredient having antioxidant activity in the extract.

In addition, the extract prepared as described above, in particular, the extract may be concentrated or removed by performing a reduced pressure filtration process or further concentrated and / or lyophilization.

Specifically, the licorice extract is crushed licorice washed with running water with a grinding mill, an extraction solvent, methylene chloride or a mixed solvent of normal hexane and ethanol, preferably a mixed solvent of normal hexane and ethanol, more preferably normal hexane and After adding normal hexane and ethanol mixed solvent having a content ratio of the mixed solvent of ethanol of 8: 1 to 10: 1 (normal hexane: ethanol), performing pressure extraction using an extractor or ultrasonic grinding extraction using an ultrasonic grinding extractor Can be performed. The extract may be prepared by performing filtration or concentrating and / or lyophilization of the obtained extract. The filtration process may remove residues left in the ethanol extract, and the water may be removed by performing the concentration and / or lyophilization.

For example, the pressure extraction is licorice, that is, a mixed solvent of normal hexane and ethanol having a content ratio of normal solvent and ethanol of 8: 1 to 10: 1 (normal hexane: ethanol) per 30 g of a ground sample, 50 mL to 500 mL , Preferably 100 mL to 300 mL, and then 5 minutes to 1 hour at 40 to 60 ° C. and 1400 psi to 1600 psi, preferably 45 to 55 ° C. and 1450 psi to 1550 psi It may be performed for 10 to 30 minutes. It can be carried out by a pressure extraction method. The extraction can be extracted by repeating three times, the extract extracted three times it can be concentrated together under reduced pressure conditions to obtain a licorice extract.

As another example, the ultrasonic pulverization extraction is licorice, that is, 100 mL to a mixed solvent of normal hexane and ethanol having a content ratio of a mixed solvent of normal hexane and ethanol per 100 g of grinding sample, 8: 1 to 10: 1 (normal hexane: ethanol). 2 L, preferably 500 mL to 1500 mL, and then 40 to 60 ° C., preferably 45 to 55 ° C. and 5 to 100 kHZ, preferably 10 to 75 kHZ, more preferably 20 to 40 kHZ Under conditions, it can be carried out by ultrasonic grinding extraction method for 10 to 30 days, preferably 14 to 21 days. The extraction can be extracted by repeating three times, the extract extracted three times it can be concentrated together under reduced pressure conditions to obtain a licorice extract.

The licorice extract is 0.4 to 0.6 parts by weight based on 100 parts by weight of licorice extract, preferably 0.45 to 0.55 parts by weight, more preferably 0.5 to 0.55 parts by weight of 7- O -methylrutheon and 0.2 to 0.4 parts by weight, preferably Preferably 0.25 to 0.35 parts by weight, more preferably 0.275 to 0.325 parts by weight of dihydrogliasperin C.

Licorice extract according to the present invention has an antioxidant activity, and has the effect of improving or preventing various diseases associated with reactive oxygen species.

The present invention also relates to an antioxidant composition comprising 7- O -methylrutheon as an active ingredient.

The 7- O -methylrutheon was measured for antioxidant activity by measuring DPPH scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (1,1'-dephenyl-2-picrylhydrazyl, DPPH). As a result, it was confirmed that there is a remarkably excellent antioxidant activity compared to the dihydrogliasperin C and the dehydroglyasperin D (Dhydroglyasperin D) that was previously confirmed to have an antioxidant activity.

An antioxidant composition comprising the 7- O -methylrutheon as an active ingredient may have antioxidant activity within the range of the antioxidant capacity of the 7- O -methylrutheon. The antioxidant composition comprising 7- O -methylrutheon as an active ingredient is a substance known to have antioxidant activity within a range of maintaining the antioxidant activity of the active ingredient 7- O -methyllutheon, for example, dihydro Gliasperin C and / or dihydrogliasperin D and the like.

The antioxidant composition of the present invention is an active ingredient licorice extract or 7- O -methylrutheon in addition to nutrients, vitamins, electrolytes, flavors, coloring agents, neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective properties Colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The components may be added to the antioxidant composition independently or in combination. The content of the additional ingredient is preferably added in the range of 0.1 to 20 parts by weight per 100 parts by weight of the licorice extract or in the range of 100 to 10,000,000 parts by weight per 100 parts by weight of 7- O -methylrutheon.

The present invention also provides an antioxidant food composition comprising licorice extract or 7- O -methylrutheon as an active ingredient. Examples of the food composition of the present invention include health food, food additives, beverages or beverage additives.

The food composition comprising the licorice extract or 7- O -methylrutheon as an active ingredient may further include appropriate carriers, excipients and diluents commonly used in the preparation thereof.

In the present specification, the term "food" means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. It is intended to include all dietary supplements, beverages, food additives and beverage additives.

Examples of the food which may include the active ingredient of the present invention include various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, the food in the present invention includes special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, ice cream, etc., natural seasonings (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages, and other health supplements. The health functional food, beverages, food additives or beverage additives may be prepared by a conventional manufacturing method.

In the present invention, a health functional food is a biological defense rhythm control, disease prevention and the like having a food group or a food composition which has added value to the food by using physical, biochemical, biotechnological techniques, etc. It means a food that is designed and processed to fully express the gymnastics function on recovery.

The dietary supplement may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of functional foods.

In the present invention, the drink is a generic term for drinking to quench thirst or to enjoy the taste and is intended to include a functional drink. The beverage is not particularly limited to other ingredients other than including the licorice extract as an active ingredient as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general drinks.

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose and other disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like, and Sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .

The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention. In addition, the composition of the present invention may further contain a pulp for the production of natural fruit juices, fruit juice drinks, vegetable drinks.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. These components can be used independently or in combination. The additive is 100 parts by weight 0 to 20 parts by weight per extract of the present invention, preferably from 0.00001 to 15 parts by weight or 7- O - methyl Lu Theon 100 parts by weight of 0 to 100,000 parts by weight of sugar, preferably from 10 to 10,000 parts by weight It may be a portion, but is not limited thereto.

In the present invention, the health functional beverage is a biological defense rhythm control or disease prevention of a beverage group or beverage composition which has added value to the beverage by using physical, biochemical and biotechnological techniques to act and express the function of the beverage for a specific purpose. Means a beverage that is designed and processed to fully express the gymnastic function related to recovery and the like.

The health functional beverage is not particularly limited to other ingredients except the licorice extract of the present invention as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose and other disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like, and xylitol Sugar alcohols such as sorbitol and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The natural carbohydrate may be included in an amount of 0 to 20 parts by weight, preferably 1 to 18 parts by weight, more preferably 5 to 12 parts by weight, per 100 parts by weight of the composition of the present invention.

In addition, in the antioxidant food composition, the amount of the licorice extract may be included as 0.00001% to 50% by weight of the total food weight, the amount of 7- O -methylrutheon is 0.00001% to 5% by weight of the total food weight When the food composition is a beverage, the amount of licorice extract is included in a ratio of 0.001 g to 50 g, preferably 0.01 g to 10 g based on 100 ml of the total volume of the food, or the 7- O − The amount of methylrutheon may be included in a ratio of 0.000001 g to 0.5 g, preferably 0.0001 g to 0.1 g, based on 100 ml of the total volume of the food, but is not limited thereto.

The antioxidant composition of the present invention is a licorice extracted from a mixed solvent of domestic licorice mixed with methylene chloride or normal hexane and ethanol 8: 1 to 10: 1 (normal hexane: ethanol), licorice grown or collected elsewhere Compared to extracts extracted from Korean extracts or other solvents, the contents of 7- O- methylluteone, Dehydroglyasperin C and Dehydroglyasperin D, which were found to be excellent in antioxidant activity and excellent in antioxidant activity, were significantly higher, and 7- O- methylluteone was remarkably high. It was confirmed that there is an excellent antioxidant activity to complete the present invention.

More specifically, domestic licorice is 7- O -methylluteone and Dehydroglyasperin C each contain about three times higher than licorice grown in Inner Mongolia, which is the highest in licorice grown or collected elsewhere in Korea. It was confirmed that there was, it was confirmed that the antioxidant activity is very excellent.

The present invention also relates to a method for preparing licorice extract having enhanced antioxidant capacity.

The method for preparing licorice extract with increased antioxidant capacity may be a method for preparing licorice extract comprising washing and grinding domestic licorice to obtain a licorice pulverized product and preparing an extract by adding an organic solvent to the licorice pulverized product. .

The domestic licorice is Glycyrrhiza It can be a glabra .

The organic solvent may be an alcohol having 1 to 5 carbon atoms, hexane, dichloromethane, methylene chloride, chloroform, ethyl acetate, ethyl ether, or a mixed solvent thereof, preferably a mixture of methylene chloride or normal hexane and ethanol It may be a solvent, more preferably a mixed solvent of normal hexane and ethanol. The content ratio of the mixed solvent of normal hexane and ethanol may be preferably 8: 1 to 10: 1 (normal hexane: ethanol) on a volume ratio basis.

The step of preparing the extract may be carried out by a pressure extraction method or an ultrasonic grinding extraction method, preferably a pressure extraction method. The pressure extraction method may be a device extraction method using a device.

The pressure extraction method may be carried out at 40 to 60 ℃ and 1400 psi to 1600 psi conditions, preferably at 45 to 55 ℃ and 1450 psi to 1550 psi conditions. In addition, the pressure extraction method may be performed for 5 minutes to 10 hours, preferably 10 minutes to 1 hour. In addition, the ultrasonic grinding extraction method may be carried out under the conditions of 40 to 60 ℃, preferably 45 to 55 ℃ and 5 to 100 kHZ, preferably 10 to 75 kHZ, more preferably 20 to 40 kHZ. In addition, the pressure extraction method may be performed for 1 day to 48 days, preferably 10 days to 30 days, more preferably 14 days to 21 days.

Licorice extract prepared by the manufacturing method is 0.4 to 0.6 parts by weight, preferably 0.45 to 0.55 parts by weight, more preferably 0.5 to 0.55 parts by weight based on 100 parts by weight of licorice extract 7- O -methylrutheon and 0.2 To 0.4 part by weight, preferably 0.25 to 0.35 part by weight, more preferably 0.275 to 0.325 part by weight of dihydro gliasperin C.

The licorice extract manufacturing method may further comprise the step of performing chromatography. The chromatography method may be a medium pressure liquid chromatography method.

The present invention also relates to a method for purifying 7- O -methylrutheon or dihydrogliasperin C from licorice.

The inventors of the present invention is an extract of domestic licorice, preferably 7- O -methylrutheon or dihydrogliasperin C, which has been found to have excellent antioxidant activity in the extract of domestic licorice extracted with a non-polar solvent, such as China or In comparison with licorice grown near Russia, the present invention was found to contain significantly more, and the optimum conditions under which 7- O -methylrutheon or dihydrogliasperin C can be purified from domestic licorice were selected. Completed.

The method for purifying 7- O -methylrutheon or dihydrogliasperin C from licorice is known as Korean licorice ( Glycyrrhiza). glabra ) to add an organic solvent and extract to prepare an extract; And purifying the extract by chromatography.

The organic solvent may be an alcohol having 1 to 5 carbon atoms, hexane, dichloromethane, methylene chloride, chloroform, ethyl acetate, ethyl ether or a mixed solvent thereof, preferably a mixed solvent of methylene chloride or normal hexane and ethanol More preferably, a mixed solvent of normal hexane and ethanol. The content ratio of the mixed solvent of normal hexane and ethanol may be 5: 1 to 15: 1, preferably 8: 1 to 10: 1 (normal hexane: ethanol) based on volume ratio.

The chromatography method may be a column chromatography method, and may be performed once to five times in consideration of yield and purity. The column chromatography method may be preferably at least one selected from the group consisting of a reduced pressure liquid chromatography method, a medium pressure liquid chromatography method, and a high performance liquid chromatography method (HPLC), and more preferably It may be at least one selected from the group consisting of a liquid chromatography method and a medium pressure liquid chromatography method.

The solvent used for the reduced pressure liquid chromatography may be at least one selected from the group consisting of alcohols having 1 to 5 carbon atoms, methylene chloride, hexane and ethyl acetate, and preferably at least one selected from the group consisting of hexane and ethyl acetate. It may be more preferably a mixed solvent of hexane and ethyl acetate. The hexane may preferably be normal hexane (n-hexane). The content of hexane and ethyl acetate may be 75:35 to 55:45 (normal hexane: ethyl acetate), preferably 70:30 to 60:40, based on volume ratio. The content range of the hexane and ethyl acetate is to adjust the polarity of the solvent in order to obtain a high purity of the purified material in high yield in consideration of the yield and purity of the material to be purified and the properties of the material to be purified.

The solvent used in the middle pressure liquid chromatography (MPLC) may be any one selected from the group consisting of water and an alcohol having 1 to 5 carbon atoms, preferably an aqueous alcohol solution of 1 to 5 carbon atoms. And, more preferably, aqueous methanol solution or aqueous ethanol solution. The concentration of the aqueous alcohol solution, that is, the alcohol content may be different depending on the type of the compound to be purified, and may be changed to give a concentration gradient while performing chromatography.

When the final result of the purification method, that is, the purified object is the 7- O -methylrutheon, the concentration gradient of the aqueous alcohol solution is 30% to 90%, preferably 35% to 80%, more preferably 40% to 70%, when the purification target is the dihydrogliasperin C concentration gradient of the aqueous alcohol solution may be 30% to 100%, preferably 45% to 100%, preferably 60% to 100%. have. The concentration gradient range of the aqueous alcohol solution is a range in which the characteristics of the substance to be purified and the polarity of the solvent are adjusted to obtain a substance to be purified having a purity of 95% or higher in high yield in consideration of the yield and purity of the substance to be purified.

The method for purifying 7- O -methylrutheon from the licorice may preferably include preparing a fraction by performing liquid chromatography under reduced pressure, and performing medium pressure liquid chromatography on the fraction.

Preferably, the method for purifying 7- O -methylrutheon from the licorice is made of Korean licorice ( Glycyrrhiza). glabra ) to a mixed solvent of methylene chloride or normal hexane and ethanol in a volume ratio of 8: 1 to 10: 1 (normal hexane: ethanol) by adding and extracting to prepare an extract; Preparing a fraction by performing reduced pressure liquid chromatography using a solvent in which n-hexane and ethyl acetate are mixed at a volume ratio of 6: 4 to 7: 3 with respect to the extract solvent; And giving a concentration gradient from 45% to 60% of the concentration of the aqueous methanol solution with respect to the fraction.

The method for purifying dihydrogliasperin C from the licorice may preferably include performing medium pressure liquid chromatography on the extract.

Preferably, the method for purifying dihydrogliasperin C from the licorice is made of domestic licorice ( Glycyrrhiza glabra ) is added to the mixed solvent of methylene chloride or normal hexane and ethanol in a volume ratio of 8: 1 to 10: 1 (normal hexane: ethanol) and extracted to prepare an extract and at 70% to the extract It may be to include the step of performing a medium pressure liquid chromatography while giving a concentration gradient of the aqueous methanol solution to 100%. The concentration gradient range of the aqueous alcohol solution is a range in which the characteristics of the substance to be purified and the polarity of the solvent are adjusted to obtain a substance to be purified having a purity of 95% or higher in high yield in consideration of the yield and purity of the substance to be purified.

The domestic licorice has a significantly higher content of 7- O -methylrutheon and dihydrogliasperin C than licorice grown in other countries except Korea, and the extract preparation method is determined by the conditions of each chromatography method. In this case, the 7- O -methylrutheon or dihydrogliasperin C can be obtained in a significantly higher yield than in the case of using another solvent or under different conditions, and 7- O obtained from the licorice. Since -methylrutheon or dihydrogliasperin C is obtained from natural products, the purification and the purification method will have a great industrial effect since it will have superior preference and stability compared with chemical synthesis.

Since the extract of the domestic licorice of the present invention with a mixed solvent of methylene chloride or normal hexane and ethanol mixed with 8: 1 to 10: 1 (normal hexane: ethanol) is not only remarkably excellent in antioxidant activity, but also using natural products, Compared with conventional chemically synthesized antioxidants, it is recognized to have an excellent effect on side effects and stability.In the case of Korean licorice, the content of 7- O- methylluteone and Dehydroglyasperin C is significantly higher than that of other plantation. In case of producing 7- O- methylluteone and Dehydroglyasperin C using domestic licorice, which is a cheap and easily available natural product in Korea, the industrial effect will be very great in terms of economy and stability.

Hereinafter, preferred embodiments of the present invention are described. However, the following examples are only preferred embodiments of the present invention, and the scope of the present invention is not limited to the following examples.

Example 1: Preparation of Licorice Extract

Example 1-1. Sample Preparation

Licorice dry powder, which is a sample to be used for licorice extract, was prepared by washing 1 kg of licorice in running water, drying at room temperature, and pulverizing using a crusher. The licorice is 11 kinds of licorice with different cultivation and harvesting areas, specifically licorice grown or harvested in Korea (Yongju), China Xinjiang, China Liaoning Jilin Province, Inner Mongolia, Uzbekistan and Kirkarstan. The licorice cultivated or collected in the Republic of Korea, Uzbekistan and Kyrgyzstan is Glycyrrhiza glabra , the licorice cultivated or harvested in Xinjiang, China is Glycyrrhiza inflata , and the licorice grown or collected in Inner Mongolia was Glycyrrhiza uralensis , Liaoning Jilin Province, China Licorice was a mixture of Glycyrrhiza inflata and Glycyrrhiza uralensis , all of the licorice was purchased from Daekwang Pharmaceutical Co., Ltd. (Chuncheon, South Korea).

Methanol, ethanol, methylene chloride and n-hexane used for licorice extract and chromatography were purchased from Burdick & Jackson (USA) in HPLC grade, and DPPH (1,1'-diphenyl-2-picrylhydrazyl) was used as Sigma- It was purchased from Aldrich Korea.

Example 1-2. Preparation of Licorice Extract

200 ml of the extraction solvent was added to 30 g of the licorice dry grinding product of Example 1-1, and the extraction solvent (ASE 300 Accelerated Solvent Extractor) was performed at 50 ° C. and 1500 psi for 20 minutes. After performing the extraction, the extracted sample was concentrated under reduced pressure at 45 ℃ using Modul spin 40 of Biotron Korea (South Korea).

The extraction solvent was extracted using water, ethanol, methylene chloride and ethanol: normal hexane (9: 1, volume ratio), respectively. At the time of extraction, instrument extraction (ASE300 ACCELERATED SOLVENT EXTRACOR) and ultrasonic grinding extraction (BRANSON Ultrasonics corporation) were performed in parallel. During extraction, 200 mL of extraction solvent was added per 30 g of the ground sample, followed by extraction for 20 minutes at a pressure of 1500 psi at 50 ° C. In ultrasonic grinding extraction, 1L of 95% ethanol was added per 100g of the ground sample. Extracted for 3 weeks. The extracted sample was concentrated under reduced pressure at 45 ℃ using Biotron corporation, Modul spin 40.

Example 2: Analysis of Licorice Extract Components by Planting or Shooting Field

The eleven licorice ethanol extracts obtained in Example 1 were analyzed using HPLC and on-line bioassay system (on-line dereplication, DPPH) as shown in FIG. 1 for antioxidant activity and component analysis. .

The HPLC used Phenomenex Luna 5 m C18 (250 × 4.6 mm), Solvent A used 2% acetic acid, Solvent B containing acetonitrile at a concentration of 20% for 0-5 minutes, 5 to 55 minutes was used with a concentration gradient of 20 to 90%, the detection wavelength (Detection wavelength) was 254 nm, the results are shown in FIG.

As shown in FIG. 1, on-line bioassay is injected with 1,1'-dephenyl-2-picrylhydrazyl (DPPH) using another pump (Pinnacle post addition pump) to the material separated through HPLC, DPPH, a stable compound as a free radical, reacts with the antioxidants passed through HPLC to determine how much of the purple color (517 nm absorption) that appears when having free radicals is measured and expressed as a negative peak. By determining the degree of free radical scavenging and the location of negative peaks, the presence of antioxidants and antioxidant activity can be measured.

In the chromatography column YMC J'Sphere ODS-H80 (250 × 4.6mm, 4㎛) was used, the results are shown in FIG. The positive peak associated with HPLC for identifying the compounds included in each extract shown in FIG. 3 is the result of analyzing the absorbance at 254 nm, in which the constituent compounds in the extract are separated from the column by time. The confirmed negative peak shows the result of analyzing the degree of absorbance decrease at 517 nm.

As shown in FIG. 2, the constituents of each licorice extract are similar in composition to the polar substances having retention time of 0 to 20 minutes, but each licorice extract in the nonpolar substance group having a retention time of 30 minutes or more. It was confirmed that the pattern was different from each other, and the activity difference of the extract according to the cultivation or harvesting site was predicted by the nonpolar substance group.

In addition, as shown in FIG. 3, antioxidants were detected in all six licorice extracts, but it was confirmed that differences in the concentration and type of the components appeared depending on the type of licorice. As confirmed in FIG. 3, the domestic licorice was found to have superior antioxidant activity compared to licorice grown or collected in other regions of Uzbekistan, Inner Mongolia, Kyrgyzstan, and Xinjiang, China. It was confirmed that the nonpolar compound had a time of 30 minutes or more.

More specifically, as shown in FIG. 3, a total of five substances showing antioxidant activity were identified. Four kinds of antioxidants were found in licorice (natural) or cultivated licorice from Inner Mongolia and cultivated in Inner Mongolia, but in case of licorice of Uzbekistan and Kyrgyzstan such as domestic licorice and paper, It was confirmed that the kind was different. In other words, the components related to antioxidants were determined by the place where licorice grew, that is, by plantation or harvesting, rather than by the type of licorice species.

On the other hand, among the substances showing the antioxidant activity, it was confirmed that the first substance (YJ-7) having a retention time of 35.4 minutes has the best antioxidant activity.

In relation to the results shown in FIG. 3, the contents of the compounds showing antioxidant activity were analyzed and shown in Tables 1 and 2 below. Table 1 is a value of measuring the width of each peak shown in Figure 3 related to the antioxidant activity, Table 2 shows the content (mg) of each component contained per 100g extract based on this.

1 (YJ-7) 2 3 4 (YJ-19) 5 (YJ-19) Republic of Korea ( G. glabra ) 527.6 65.3 298.2 114.8 Uzbekistan ( G. glabra ) 289.5 125.9 Xinjiang ( G. inflata ) 137.2 Kyrgyzstan ( G. glabra ) 35.9 67.4 Inner Mongolia ( G. uralensis ) 77.8 35.6 120.1 115.3 Inner Mongolia (cultivation) ( G. uralensis ) 228.2 127.9 133.5 96.8

As shown in Table 1 and Table 2, in the case of domestic licorice, it was confirmed that the antioxidant activity is remarkably superior to other licorice, the content of YJ-7 is confirmed that the best antioxidant activity is the other licorice in the case of domestic licorice It is about 2.5 to 18 times as compared to the licorice cultivated in Inner Mongolia, which has the highest content among licorice except domestic licorice. It is confirmed that the content of YJ-19 is found to be excellent in antioxidant activity. Licorice was about 2.5 to 3 times higher than other licorice, and it was confirmed that it exhibited about 2.5 times the content of licorice grown in Inner Mongolia, which has the highest content among licorice except domestic licorice. In addition, YJ-20 also showed the same activity tendency as YJ-19, it was confirmed that the content is similar.

Therefore, it was confirmed from the above results that the domestic licorice is very excellent in antibacterial activity compared to the licorice grown or collected in other regions except Korea.

Example  3: Analysis of Licorice Extract Components According to Extraction Solvents

In order to measure the antioxidant activity according to the extraction solvent of the domestic licorice extract, which was found to have the best antioxidant activity in Example 2, water, ethanol, methylene chloride and ethanol: normal hexane (9: 1, For licorice extract extracted by volume ratio), HPLC analysis was performed under the conditions of Example 2, and the results are shown in FIG. 4.

As shown in FIG. 4, the extraction solvent was identified to be different in the form of the water, ethanol, methylene chloride and ethanol: normal hexane (9: 1, volume ratio) extracted depending on the polarity of the solvent.

Specifically, the water extract compared to the other solvents, almost non-polar material is extracted, only the polar material (Rt, 30min or less) was extracted, the ethanol extract was extracted both polar and non-polar material. On the other hand, methylene chloride and ethanol: normal hexane (9: 1, volume ratio), which are nonpolar solvents, were found to extract almost similar materials.

In addition, the extract of the non-polar solvent methylene chloride and ethanol: normal hexane (9: 1, volume ratio) was confirmed to have the best antioxidant activity, the preferred solvent for preparing the extract is a mixed solvent of methylene chloride and ethanol and normal hexane Afterwards, it was determined that the studies on the substance having antioxidant activity should be performed on the nonpolar substance of the domestic licorice extract.

Example  4: from domestic licorice extract 7- O -methylluteone, Dehydroglyasperin  C and Dehydroglyasperin  Establish production conditions of D

4-1. Identification of Major Components of Korean Licorice Extract

In Example 3, the substance contained in the extract of ethanol: normal hexane (9: 1, volume ratio), which was found to have the best antioxidant activity, was subjected to the conditions of Example 2, specifically, Phenomenex Luna 5 m C18 (250). × 4.6 mm), Solvent A uses 2% acetic acid, Solvent B contains acetonitrile at a concentration of 20% for 0 to 5 minutes and 20 to 90% for 5 to 55 minutes. It was used while giving a concentration gradient, the detection wavelength (Detection wavelength) was determined to 254 nm, HPLC was performed, the results are shown in Figure 5a.

5A was added to the compound identified in FIG. 5A, and the YJ-7 (No. 7 of FIG. 5A) and YJ-19 (19 of FIG. 5A), which were found to be excellent in the antioxidant activity and contained in large amounts in the domestic licorice extract, In order to obtain YJ-20 (No. 20 in FIG. 5A) and each compound, middle pressure liquid chromatography (MPLC) was performed while giving a concentration gradient with 20% to 100% aqueous methanol solution. Shown in

As shown in FIG. 5B, it was confirmed that the compound of FIG. 5A could be obtained separately using the concentration gradient of the solvent of MPLC.

In order to determine the structures of YJ-7 and YJ-19 and YJ-20, which were found to have excellent antioxidant activity, 1H-, 13C-NMR, 1H-1H COSY, DEPT, HBQC, and HMBC spectroscopic methods were performed on each compound. The results were confirmed and the structure was determined based on these results.

Each result is as follows.

In the case of YJ-7 it was determined to 7- O -methylluteone having the following formula (1).

2 ', 4', 5-Trihydroxy-7-methoxy-6-prenylisoflavone, Molecular formula: C ₂₁ H ₂₀ O ₆ , EI-MS 368, 1H-NMR (600MHz, DMSO-d6) δ: 1.62 (3H, s , H-5 "), 1.73 (3H, s, H-4"), 3.26 (2H, d, J = 6.87 Hz, H-1 "), 3.90 (3H, s, 7-OCH ₃ ), 5.14 ( 1H, t-like, H-2 "), 6.30 (1H, dd, J = 8.24, 2.3, H-5 '), 6.40 (1H, s, J = 2.3, H-3'), 6.69 (1H, s, H-8), 7.00 (1H, d, J = 8.22, H-6 '), 8.24 (1H, s, H-2), 9.33 (1H, s, 2'-OH), 9.41 (1H, s, 4'-OH), 13 C-NMR (150 MHz, DMSO-d6) δ: 18.06 (C-4 "), 21.44 (C-1"), 25.90 (C-5 "), 56.75 (-OCH ₃ ) , 90.51 (C-8), 103.15 (C-3 '), 105.77 (C-4a), 106.72 (C-5'), 109.09 (C-1 '), 112.04 (C-6), 121.17 (C- 3), 122.40 (C-2 "), 131.40 (C-3"), 132.62 (C-6 '), 155.89 (C-2), 156.36 (C-8a), 156.86 C-2'), 158.18 ( C-5), 159.09 (C-4 '), 163.19 (C-7), 181.08 (C-4)

In the case of YJ-19 it was determined as Dehydroglyasperin C having the following formula (2) by the following results.

Molecular formula: C ₂₁ H ₂₂ O ₅ , EI-MS 354, 1H-NMR (600MHz, DMSO-d6) δ: 1.62 (3H, s, H-5 "), 1.7 (3H, s, H-4") , 3.14 (2H, d, J = 6.54, H-1 "), 3.67 (3H, s, 5-OCH ₃ ), 4.83 (2H, s, 2-CH ₂ ), 5.13 (1H, t-like, H -2 "), 6.14 (1H, s, H-8), 6.25 (1H, dd, J = 8.3, 2.4, H-5 '), 6.32 (1H, s, J = 2.4, H-3'), 6.64 (1H, s, H-4), 7.04 (1H, d, J = 8.3, H-6 '), 9.39 (1H, s, 4-OH), 9.54 (1H, s, 7-OH), 9.60 (1H, s, 2'-OH), 13 C-NMR (150 MHz, DMSO-d6) δ: 18.12 (C-4 "), 22.61 (C-1"), 25.94 (C-5 "), 62.10 (C -5-OCH ₃ ), 67.73 (C-2CH ₂ ), 98.89 (C-8), 103.18 (C-3 '), 107.30 (C-5'), 109.21 (C-4a), 114.36 (C-6 ), 115.06 (C-4), 117.02 (C-1 '), 124.37 (C-2 "), 127.97 (C-3), 129.12 (C-6'), 129.95 (C-3"), 152.74 ( C-8a), 155.38 (C-5), 156.24 (C-7), 156.57 (C-2 '), 158.46 (C-4')

In the case of YJ-20 it was determined as Dehydroglyasperin D having the following formula (2).

Molecular formula: C ₂₂ H ₂₄ O ₅ , EI-MS 368, 1H-NMR (600MHz, DMSO-d6) δ: 1.68 (3H, s, H-5 "), 1.76 (3H, s, H-4") , 3.23 (2H, d, J = 6.85, H-1 "), 3.73 (3H, s, 5-OCH ₃ ), 3.8 (3H, s, 7-OCH ₃ ), 4.95 (1H, s, 2-CH2 ), 5.14 (1H, t-like, H-2 "), 6.31 (1H, dd, J = 8.37, 2.4, H-5 '), 6.38 (1H, s, H-8), 6.4 (1H, s , J = 2.4, H-3 '), 6.74 (1H, s, H-4), 7.12 (1H, d, J = 8.37, H-6'), 9.47 (1H, s, H-4'-OH ), 9.69 (1H, s, H-2'-OH), 13 C-NMR (150 MHz, DMSO-d6) δ: 18.08 (C-4 "), 22.53 (C-1"), 25.94 (C-5 " ), 56.19 (C-7-OCH ₃ ), 62.22 (C-5-OCH ₃ ), 67.80 (C-2CH ₂ ), 95.75 (C-8), 103.25 (C-3 '), 107.36 (C-5 '), 114.67 (C-4), 115.53 (C-6), 116.71 (C-1'), 124.00 (C-2 "), 128.71 (C-3), 129.20 (C-6 '), 130.40 ( C-3 "), 153.16 (C-8a), 154.88 (C-5), 156.53 (C-2 '), 157.99 (C-7), 158.45 (C-4')

YJ-19 identified as Dehydroglyasperin C and YJ-20 identified as Dehydroglyasperin D were reported to have excellent antioxidant activity, but the antioxidant capacity of YJ-7 identified as 7- O -methylluteone was first confirmed in this experiment. It became.

4-2. 7- O -methylluteone, Dehydroglyasperin  C and Dehydroglyasperin  Establish conditions for mass production of d

The 7- O - methyl MPLC result of Lu Theon, in order to improve the purity and yield of 7- O -methylluteone identified the need for execution of additional chromatography.

Decompression liquid chromatography (VLC) was determined by a method suitable for the above additional chromatography, and it was determined to use a mixture of normal hexane and ethyl acetate as its solvent.

In order to confirm the optimum mixing ratio of the normal hexane and ethyl acetate, the pressure ratio of the liquid chromatography was performed by changing the mixing ratio, and the result of the chromatography and 7- O -methylrutheon were thin-film chromatography (TLC). Compared.

The comparison results are shown in FIG. 6A, and a band similar to 7- O -methylrutheon was found to be the most suitable in a solvent in which normal hexane and ethyl acetate were mixed (65:35, volume ratio). In addition, as a result of performing the optimum conditions of MPLC for the result of the thin layer chromatography, using the solvent of the MPLC while giving a concentration gradient from 45% to 60% methanol aqueous solution to obtain the best yield and purity It was confirmed that.

Based on these experiments, 7- O -methylrutheon was prepared under reduced pressure liquid chromatography using a solvent obtained by mixing a domestic licorice extract with normal hexane and ethyl acetate (65:35, volume ratio) to prepare a fraction. Fractions were purified by the method of performing MPLC giving a concentration gradient from 45% to 60% of the methanol aqueous solution.

On the other hand, dihydrogliasperin C performed the optimum conditions of MPLC for the domestic licorice extract, it is the best yield and purity to use the solvent of the MPLC while giving a concentration gradient from 70% to 100% methanol solution It was confirmed to obtain. The result of MPLC performed while giving a concentration gradient of the aqueous methanol solution from 70% to 100% is shown in FIG. 6B, and as shown in FIG. 6B, a section in which the same compound can be obtained is dihydrogliasperin. In the case of YJ-19 corresponding to C and YJ-20 corresponding to dihydrogliasperin D, a wide range was confirmed, and the concentration gradient condition was identified as the most preferable condition from 70% to 100%.

For a method for purifying the methyl base Theon, 7- O - - a result of the purification under the above conditions, was to be about 98% pure, purified compounds, the yield of methyl 7- O Lu Theon is 0.3% In the case of the purification method of the dihydrogliasperin C, the yield of the dihydrogliasperin C was confirmed to be about 0.15%, and purified in the same manner as the purification method of the dihydrogliasperin C , The yield of dihydrogliasperin D was found to be about 0.05%.

Example  5: Preparation of health functional food containing domestic licorice extract

Health foods containing licorice extract can be prepared by mixing the raw materials of Table 3 below.

ingredient content Licorice extract powder 1000 mg Vitamin mixtures Quantity Vitamin A Acetate 70 μg Vitamin E  1.0 mg Vitamin B1 0.13 mg Vitamin B2  0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 μg Vitamin c 10 mg Biotin 10 μg Nicotinic acid amide 1.7 mg Folic acid 50 μg Calcium Pantothenate 0.5 mg Mineral mixture Quantity Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium phosphate monobasic 15 mg Dicalcium Phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio 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.

1 is a schematic diagram showing a process of performing an on-line bioassay according to an embodiment of the present invention.

Figure 2 is a diagram showing the HPLC results of licorice ethanol extract according to the origin, according to an embodiment of the present invention.

Figure 3 is a diagram showing the results of performing an On-line DPPH-HPLC bioassay confirming the components having an antioxidant capacity using the HPLC results and DPPH of licorice ethanol extract according to the embodiment of the present invention.

Figure 4 is a diagram showing the HPLC results of the licorice extract according to the extraction solvent, according to an embodiment of the present invention.

Figure 5a is a diagram showing the HPLC results of the Korean licorice extract, according to an embodiment of the present invention.

Figure 5b is a chart showing the HPLC results of the independent Korean licorice extract by fractions according to an embodiment of the present invention.

FIG. 6A is a graph showing thin-film chromatography (TLC) results for each solvent under reduced pressure chromatography to confirm fractionation conditions of YJ-7 according to one embodiment of the present invention.

6B is a chart showing MPLC results for YJ-19 and YJ-20, according to an embodiment of the present invention.

7 is a view showing the chemical structure of dihydrogliasperin C, according to an embodiment of the present invention.

Claims (10)

An antioxidant composition comprising extract as an active ingredient extract of at least one selected from the group consisting of alcohols of 1 to 5 carbon atoms, hexane, dichloromethane, methylene chloride, chloroform, ethyl acetate, and ethyl ether as an extraction solvent. The method of claim 1, The extraction solvent is an antioxidant composition of methylene chloride or a mixed solvent in which normal hexane and ethanol are mixed with 8: 1 to 10: 1 (normal hexane: ethanol). The method of claim 1, The licorice is Glycyrrhiza glabra antioxidant composition. The method according to any one of claims 1 to 3, Which comprises a base Theon methyl (7- O -methylluteone) and 0.2 to 0.4 parts by weight of dihydro article ria peorin C (Dehydroglyasperin C) - the extract is Licorice extract 100 parts by weight of 0.4 to 0.6 parts by weight of the 7- O reference Antioxidant composition. An antioxidant food composition comprising an extract extracted by using a mixed solvent obtained by mixing methylene chloride or normal hexane with ethanol in an amount of 8: 1 to 10: 1 (normal hexane: ethanol) as an extraction solvent. The method of claim 5, Which comprises a base Theon methyl (7- O -methylluteone) and 0.2 to 0.4 parts by weight of dihydro article ria peorin C (Dehydroglyasperin C) - the extract is Licorice extract 100 parts by weight of 0.4 to 0.6 parts by weight of the 7- O reference Antioxidant food composition. 7- O - methyl Lu Theon (7- O -methylluteone) an antioxidant composition comprising, as an active ingredient. 7- O - methyl Lu Theon (7- O -methylluteone) an antioxidant food composition comprising, as an active ingredient. Domestic licorice ( Glycyrrhiza glabra ) to a mixed solvent of methylene chloride or normal hexane and ethanol in a volume ratio of 8: 1 to 10: 1 (normal hexane: ethanol) by adding and extracting to prepare an extract; Preparing a fraction by performing vacuum liquid chromatography (VLC) on the extract solvent by using a solvent in which n-hexane and ethyl acetate are mixed at a volume ratio of 6: 4 to 7: 3; And Performing a middle pressure liquid chromatography (MPLC) while giving a concentration gradient of the aqueous methanol solution from 45% to 60% for the fractions Method for purifying the methyl base Theon (7- O -methylluteone) - 7- O from licorice. Preparing an extract by adding and extracting a mixed solvent obtained by mixing methylene chloride or normal hexane and ethanol in a volume ratio of 8: 1 to 10: 1 (normal hexane: ethanol) to domestic licorice ( Glycyrrhiza glabra ); and Performing a middle pressure liquid chromatography (MPLC) while giving a concentration gradient of the aqueous methanol solution from 70% to 100% for the extract A method for purifying dehydroglyasperin C from licorice.

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