KR20140137640A - Complex extract of Quercus variabilis fractions and composition for treating or preventing diabetes comprising the same - Google Patents

Abstract

The present invention relates to a complex extract of Quercus variabilis fractions and a composition comprising the same as an active ingredient for treating or preventing diabetes. The complex extract of Quercus variabilis fractions is a mixture of certain fractions of the Quercus variabilis fractions with a predetermined ratio, has excellent anti-diabetes activities, and is a safe substance which is derived from natural tree resources; thereby can be useful for developing a medicine for treating or preventing diabetes or a functional food for alleviating diabetes.

Description

[0001] The present invention relates to a complex extract of Quercus variabilis fractions and compositions for diabetes,

The present invention relates to a complex extract of oak-leaf and a composition for treating or preventing diabetes comprising the extract as an active ingredient.

Diabetes is caused by insulin secretion degradation due to the destruction or dysfunction of the pancreatic? -Cells that secrete insulin in vivo, and by the rejection of insulin receptor in the tissue by genetic factors or obesity, glucose is excreted through the urine The metabolic diseases are characterized by hyperglycemia and hyperglycemia.

When diabetes is induced, the secretion of insulin and glucagon is disturbed, resulting in abnormal metabolic control of not only carbohydrate but also protein and lipid metabolism, resulting in various metabolic diseases. When diabetic symptoms persist for a long time, And the occurrence of many complications in circulating machines and the like is becoming a problem.

The most important goal in the treatment of diabetes is to regulate blood sugar levels as close to normal as possible. Therapeutic methods include pharmacotherapy, diet and exercise therapy. Currently, oral hypoglycemic agents used in patients with type 1 and type 2 diabetes include α-glucosidase inhibitors, sulfonylurea agents and biguanide agents, and α- Glucosidase inhibitors show the therapeutic effect of diabetes by reducing the postprandial blood glucose and elevation of blood insulin by delaying the digestion and absorption of carbohydrates in the ingested diets. Alpha-glucosidase inhibitors promote the secretion in the small intestine of glucagon-like peptide-1, which does not cause hyperinsulinemia or hypoglycemia, promotes insulin secretion and inhibits glucagon secretion It has advantages.

Acarbose, voglibose and miglitol are currently used in clinical practice. However, long-term administration of α-glycosidase inhibitor may cause side effects such as abdominal bloating and vomiting diarrhea There is a problem that its use is limited.

On the other hand, Korean Patent Registration No. 0417287 discloses a composition for treating diabetes containing a lower alcohol-insoluble fraction of Hovenia dulcinus or a polysaccharide substance separated therefrom. However, there is still a need to develop an antidiabetic extract derived from a natural wood source.

Accordingly, it is an object of the present invention to accomplish the present invention by confirming that a complex extract of a specific fraction of oak has excellent antidiabetic activity while conducting research on an antidiabetic extract derived from a natural woody resource.

Accordingly, an object of the present invention is to provide an extract of oak-leaf mixture having antidiabetic activity.

Another object of the present invention is to provide a composition for the treatment or prevention of diabetes comprising an extract of the oak-leaf mixture as an active ingredient.

In order to achieve the above object, the present invention relates to a method for treating oak, which comprises 70 to 90% by weight of a volatile fraction obtained from an oak and 10 to 30% by weight of a non-volatile fraction obtained from an oak, wherein the volatile fraction and non- step; Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water; Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And concentrating the liquid extract to produce a volatile fraction and a non-volatile fraction, respectively.

The present invention also provides a pharmaceutical composition for the treatment or prevention of diabetes, comprising as an active ingredient an extract of oak fraction comprising 70 to 90% by weight of a volatile fraction obtained from oak and 10 to 30% by weight of a non-volatile fraction obtained from oak, The volatile fraction and the non-volatile fraction treating the oak at a high temperature and a high pressure; Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water; Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And concentrating the liquid extract to prepare a volatile fraction and a non-volatile fraction, respectively, to provide a pharmaceutical composition for the treatment or prevention of diabetes.

The present invention also provides a functional food for diabetes mellitus improvement comprising an extract of the oak-leaf mixture containing 70 to 90% by weight of a volatile fraction obtained from oak and 10 to 30% by weight of a non-volatile fraction obtained from oak as an active ingredient, And a non-volatile fraction treating the oak at a high temperature and a high pressure; Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water; Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And concentrating the liquid extract to produce a volatile fraction and a non-volatile fraction, respectively, to provide a diabetic functional food for improving diabetes.

More specifically, the volatile fraction and the nonvolatile fraction are subjected to a high-temperature and high-pressure treatment at a temperature of 190 to 250 ° C at a pressure of 20 to 30 kgf / cm ² for 2 to 30 minutes; Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and subjecting it to hydrothermal extraction at a temperature of 55 to 65 ° C for 1 to 6 hours; Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And concentrating the liquid extract at 75 to 85 캜 under vacuum for 1 to 3 hours to prepare a volatile fraction and a non-volatile fraction, respectively.

The oak-fraction complex extract according to the present invention is obtained by mixing specific fractions of oak fractions at a specific weight ratio and exhibits excellent antidiabetic activity, and is a safe substance derived from natural wood resources. Therefore, the extract of oak- And can be very useful for the development of functional foods for diabetes mellitus improvement.

1 is a process diagram for obtaining an oak-fraction complex extract according to the present invention.

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

The present invention relates to a method for treating oak, comprising the steps of: 70 to 90% by weight of a volatile fraction obtained from an oak; and 10 to 30% by weight of a non-volatile fraction obtained from an oak, wherein the volatile fraction and the non- Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water; Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And concentrating the liquid extract to produce a volatile fraction and a non-volatile fraction, respectively.

Wherein said volatile fraction and a non-volatile fraction is treated for 190 to the oak at a temperature of 250 ℃ at a pressure of 20 to 30 kgf / cm ^{2 2} to 30 minutes high temperature and high pressure; Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and subjecting it to hydrothermal extraction at a temperature of 55 to 65 ° C for 1 to 6 hours; Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And concentrating the liquid extract at 75 to 85 캜 under vacuum for 1 to 3 hours to obtain a volatile fraction and a non-volatile fraction, respectively.

The extract of the above-mentioned oak fraction exhibits an excellent antidiabetic activity in the above range, and exhibits the most excellent antidiabetic activity especially when the volatile fraction obtained from oak is 70 wt% and the non-volatile fraction obtained from oak is 30 wt%.

The present invention also provides a pharmaceutical composition for the treatment or prevention of diabetes, comprising the above-mentioned complex extract of oak fraction as an active ingredient.

The pharmaceutical composition may contain a pharmaceutically acceptable carrier in addition to the complex extract of the oak fraction. Such pharmaceutically acceptable carriers are those conventionally used in pharmaceutical preparations, and include lactose, dextrose, sucrose, sorbitol, But are not limited to, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, Talc, magnesium stearate, mineral oil, and the like, but is not limited thereto. In addition, the pharmaceutical composition may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. as an additive.

The pharmaceutical composition may be administered according to the severity of diabetes mellitus. Usually, local administration is preferred. The dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the severity of the disease, the age, sex, and weight of the patient, and may be administered once to several times per day.

The pharmaceutical composition may be prepared in unit dose form by formulating it with a pharmaceutically acceptable carrier and / or excipient, or may be prepared by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions, or may be in the form of elixirs, excipients, powders, granules, tablets, alerts, lotions, ointments and the like.

The present invention also provides a functional food for diabetes mellitus containing the above-mentioned complex extract of oak fraction as an active ingredient.

The functional food may be a gum, a vitamin complex, a health supplement, a nutritional supplement food, a functional beverage, or the like. In addition to the active ingredient oak extract compound extract, there may be mentioned glucose, monosaccharide such as fructose, maltose, Disaccharides such as sucrose or polysaccharides such as dextrin and cyclodextrin may be added and sugar alcohols such as xylitol, sorbitol and erythritol may be added and sweeteners such as taurine and stevia extract may also be added have.

For the purpose of improving the diabetes, when the complex extract of oak is added to the food, it is generally added in an amount of 1 to 20% by weight of the total food, and when added to the beverage, 0.1 to 100 mL, Preferably at a content ratio of 10 to 100 mL.

On the other hand, the extract of the oak-fraction complex according to the present invention is a natural tree-derived material and has little toxicity and side effects, so that it can be safely used for prolonged use even for prophylactic purposes.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the present invention is not limited by these examples.

Example 1: Production of oak-fraction extract

The extracted material was Quercus variabilis wood chips (5 cm x 5 cm x 0.5 cm) treated with steam at 250 DEG C for 15 minutes at a pressure of 25 kgf / cm < ² >. To 10 kg of the wood chips, 200 L of distilled water And the mixture was subjected to hot water extraction at 60 ° C for 3 hours. The extract was centrifuged at 3000 rpm for 30 minutes, and the supernatant was separated and filtered through a 0.45 μm membrane filter (Sartorius ^® , Germany) to obtain a first yellow liquid extract (Fraction 1). Then, the first liquid extract was concentrated using a concentrator at 80 ° C for 2 hours to obtain a transparent volatile fraction (Fraction 2) and a brown nonvolatile fraction (Fraction 3). Then, 4000 mL of ethanol was added to the residue after the centrifugation, and the mixture was centrifuged at 3000 rpm for 5 minutes. The supernatant was separated to obtain a purple second liquid extract (Fraction 4).

≪ Example 2 > Analysis of components of oak-fraction extract

1. Volatile Compound Analysis

1 mL of each fraction prepared in Example 1 was added to a 10 mL vial for head space, and the mixture was heated for 15 minutes and analyzed by GC / MS with a head space autosampler (Varian, USA).

At this time, GC / MS analysis conditions are as follows.

Column: DB-5MS (30 m x 0.25 mm i.d., 0.25 m)

- Injector temperature: 250 ℃

Carrier gas: helium, 0.8 mL / min

- Split ratio = 1: 20

- Oven temperature: maintained at 40 캜 for 5 minutes, then increased to 250 캜 at a rate of 5 캜 / min, maintained at 250 캜 for 10 minutes

- GC-MS library: National Institute of Standards and Technology (NIST)

As a result, 21 kinds of substances were present in the first liquid extract (Fraction 1) (Table 1), 32.4% in the case of Fetradecane, 21.7% in the acrylic acid and 18.8% in the acetic acid, Compared to other materials, these three materials showed a relatively high content. In addition, the content of hexadecane was 4.9%, that of furan-2-carboxaldehyde was 4.3% and that of 1,2,3-trichloroheptachlorobutane was 2.4%, and a small amount of the compound at a level of 0.5% to 1.8% Respectively. At this time, RT means retention time (min).

No RT ^a compound The content, %  One  2.232 Acetic acid C ₂ H ₄ O ₂ 18.8  2  7.002 Furfural C ₅ H ₄ O ₂ 4.3  3  7.579 2-Furanmethanol C ₅ H ₆ O ₂ -  4  9.882 Ocimenyl acetate C ₁₂ H ₁₈ O ₂ 0.7  5 10.002 Linalyl propionate C ₁₃ H ₂₂ O ₂ 1.2  6 10.103 Methanesulfonic acid CH ₄ O ₃ S 1.0  7 11.457 Decane C ₁₀ H ₂₂ 0.7  8 12.545 Butane C ₄ H ₁₀ 1.0  9 13.83 Endo-borneol C ₁₀ H ₁₈ O 0.7 10 14.133 6-Dodecanone C ₁₂ H ₂₄ O 0.6 11 14.234 Benzoic acid C ₇ H ₆ O ₂ 0.5 12 15.640 Isobornyl thiocyanoacetate C ₁₃ H ₁₉ NO ₂ S 0.6 13 16.534 2-Hexadecanol C ₁₆ H ₃₄ O 0.9 14 16.724 1,2,3-trichloroheptachlorobutane C ₄ Cl ₃ F ₇ 2.4 15 16.886 Bornyl ester of acrylic acid C ₁₃ H ₂₀ O ₂ 21.7 16 17.059 Ethanol C ₂ H ₆ O 1.8 17 17.251 Pentadecane C ₁₅ H ₃₂ 1.6 18 18.653 Octadecane C ₁₈ H ₃₈ 1.3 19 18.857 Hexadecane C ₁₆ H ₃₂ 4.9 20 19.177 Tetradecane C ₁₄ H ₃₀ 32.4 21 19.563 Heptadecane C ₁₇ H ₃₆ - 22 20.498 1-Heptacosanol C ₂₇ H ₅₆ O 0.7 23 22.723 3,4-Dimethoxy-dl-phenylalanine C ₁₁ H ₁₅ NO ₄ 2.1 Sum 100

In the volatile fraction (Fraction 2), there were 14 kinds of substances (Table 2), and the highest percentage of furfural was 65.2%. In addition, the contents of acrylic ester and acrylic acid were 9.9% and 9.5%, respectively, and small amounts of compounds were detected at levels of 0.3% to 5.0%.

No RT ^a compound The content, %  One  2.232 Acetic acid C ₂ H ₄ O ₂ 1.2  2  7.002 Furfural C ₅ H ₄ O ₂ 65.2  3  7.579 2-Furanmethanol C ₅ H ₆ O ₂ -  4  9.882 Ocimenyl acetate C ₁₂ H ₁₈ O ₂ -  5 10.002 Linalyl propionate C ₁₃ H ₂₂ O ₂ 5.0  6 10.103 Methanesulfonic acid CH ₄ O ₃ S -  7 11.457 Decane C ₁₀ H ₂₂ -  8 12.545 Butane C ₄ H ₁₀ 0.5  9 13.830 Endo-borneol C ₁₀ H ₁₈ O - 10 14.133 6-Dodecanone C ₁₂ H ₂₄ O - 11 14.234 Benzoic acid C ₇ H ₆ O ₂ - 12 15.640 Isobornyl thiocyanoacetate C ₁₃ H ₁₉ NO ₂ S - 13 16.534 2-Hexadecanol C ₁₆ H ₃₄ O 0.6 14 16.724 1,2,3-trichloroheptachlorobutane C ₄ Cl ₃ F ₇ 1.2 15 16.886 Bornyl ester of acrylic acid C ₁₃ H ₂₀ O ₂ 9.9 16 17.059 Ethanol C ₂ H ₆ O 1.1 17 17.251 Pentadecane C ₁₅ H ₃₂ 0.8 18 18.653 Octadecane C ₁₈ H ₃₈ 0.6 19 18.857 Hexadecane C ₁₆ H ₃₂ 2.4 20 19.177 Tetradecane C ₁₄ H ₃₀ 9.5 21 19.563 Heptadecane C ₁₇ H ₃₆ - 22 20.498 1-Heptacosanol C ₂₇ H ₅₆ O 0.3 23 22.723 3,4-Dimethoxy-dl-phenylalanine C ₁₁ H ₁₅ NO ₄ 1.7 Sum 100

In the non-volatile fraction (Fraction 3), 21 kinds of substances are present (Table 3), and the content of the tetradecane is 24.8%, the ester of acrylic acid is 23.3%, and the content of acetic acid is 21.5% And showed a relatively high content. In addition, furan-2-carboxaldehyde showed a content of 9.1% and a small amount of compound was detected at a level of 0.3% to 3.5%.

No RT ^a compound The content, %  One  2.232 Acetic acid C ₂ H ₄ O ₂ 21.5  2  7.002 Furfural C ₅ H ₄ O ₂ 9.1  3  7.579 2-Furanmethanol C ₅ H ₆ O ₂ -  4  9.882 Ocimenyl acetate C ₁₂ H ₁₈ O ₂ 2.0  5 10.002 Linalyl propionate C ₁₃ H ₂₂ O ₂ 2.0  6 10.103 Methanesulfonic acid CH ₄ O ₃ S 2.6  7 11.457 Decane C ₁₀ H ₂₂ 0.6  8 12.545 Butane C ₄ H ₁₀ 1.0  9 13.83 Endo-borneol C ₁₀ H ₁₈ O 0.5 10 14.133 6-Dodecanone C ₁₂ H ₂₄ O 0.4 11 14.234 Benzoic acid C ₇ H ₆ O ₂ 0.4 12 15.64 Isobornyl thiocyanoacetate C ₁₃ H ₁₉ NO ₂ S 0.6 13 16.534 2-Hexadecanol C ₁₆ H ₃₄ O 0.7 14 16.724 1,2,3-trichloroheptachlorobutane C ₄ Cl ₃ F ₇ 3.5 15 16.886 Bornyl ester of acrylic acid C ₁₃ H ₂₀ O ₂ 23.3 16 17.059 Ethanol C ₂ H ₆ O 1.0 17 17.251 Pentadecane C ₁₅ H ₃₂ 0.3 18 18.653 Octadecane C ₁₈ H ₃₈ 1.7 19 18.857 Hexadecane C ₁₆ H ₃₂ 2.3 20 19.177 Tetradecane C ₁₄ H ₃₀ 24.8 21 19.563 Heptadecane C ₁₇ H ₃₆ - 22 20.498 1-Heptacosanol C ₂₇ H ₅₆ O 0.7 23 22.723 3,4-Dimethoxy-dl-phenylalanine C ₁₁ H ₁₅ NO ₄ 1.0 Sum 100

 There are 17 kinds of substances in the second liquid extract (Fraction 4) (Table 4), and ethanol content is 15.17% and methyl 2-chloro-2-methyl propionate content is 13.4% The material showed a relatively high content. In addition, the content of E-1-methoxy-7-methyl-1,6-octadiene was 8.58%, and a small amount of compound was detected at a level of 0.3% to 3.5%.

No RT ^a compound The content, %  One  1.799 Ethyl 2-chloropropionate C ₅ H ₉ ClO ₂  13.4  2  2.324 Ethanol CH ₃ CH ₂ OH  15.2  3  2.488 Geraniol C ₁₀ H ₁₈ O   8.6  4  3.484 Methanamine, N
- (1-methylbutylidene) - C ₆ H ₁₃ N   6.8  5  4.248 4-fluoropiperidine C ₅ H ₁₀ FN   4.4  6  4.751 Ibuprofen C ₁₃ H ₁₈ O ₂   4.6  7  5.537 4-Hydroxy-2,2,6,6-tetramethylpiperidinol C ₉ H ₁₉ NO   2.9  8  5.923 Decanal, O-methyloxime C ₁₁ H ₂₃ NO   7.1  9  7.142 3-Hydroxy-3-phenylvaleric acid C ₁₁ H ₁₄ O ₃   5.7 10  7.500 alpha-pinene C ₁₀ H ₁₆   4.2 11 10.228 Phenyl isobutyrate C ₁₀ H ₁₂ O ₂   3.5 12 13.872 Thiophene-2-acetic acid, cyclobutylester C ₁₂ H ₁₆ O ₂ S   3.0 13 14.723 Divinylmethyl (chloromethyl) silane C ₆ H ₁₁ ClSi   3.8 14 15.932 3-bicyclo [2.2.1] heptanyloxy-dimethyl-prop-2-enylsilane C ₁₂ H ₂₂ OSi   3.5 15 16.083 1- (3,4-Methylenedioxybenzylidene) semicarbazide C ₉ H ₉ N ₃ O ₃   3.1 16 23.609 2 - [(3R) -quinuclidin-3-yl] -3H-benzo [g] isoindol-1- C ₁₉ H ₂₀ N ₂ O   4.9 17 25.080 6-Oxtadecanone C ₁₈ H ₃₆ O   5.5 Sum 100

 Therefore, substances commonly contained in the first liquid extract (Fraction 1), volatile fraction (Fraction 2) and non-volatile fraction (Fraction 3) include tetradecane and acrylic acid vinyl ester, , Which is used as a raw material for production of paraffin wax, and the highest content was 32.4% in the first liquid extract (Fraction 1). The acrylic acid vinyl ester had the highest content of acrylic acid derivative (23.3%) in the nonvolatile fraction (Fraction 3).

Acetic acid contained in the first liquid extract (Fraction 1) and the nonvolatile fraction (Fraction 3) was one of important carboxylic acids having sterilizing ability and showed a content of 21.5% in the nonvolatile fraction (Fraction 3). Perfural is a type of heterocyclic aldehyde in the form of an oil-like liquid with a special odor. It is the compound with the highest content of 65.2% in the volatile fraction (Fraction 2). It is used for nylon synthesis and insecticide. The second liquid extract (Fraction 4) was obtained by extracting the oak from the blast-treated hot water and then extracting the residue again with ethanol. The ethanol content of the extract was 15.2% and the content of 8.6% Lanolin is a kind of mono terpenoid, which is used as a perfume in cosmetic production.

2. Solid content analysis

1 mL of each fraction prepared in Example 1 was placed in a full-length container and dried in a dryer at 105 ° C for 24 hours. After drying, the container was allowed to cool and then weighed and substituted into the following equation to calculate the solid content.

[Equation 1]

Solid content (g / mL) = [total container weight (g) + sample weight (g)] - total container weight (g)

As a result, the solid content of the non-volatile fraction (Fraction 3) was the highest at 12.4 g / L and the solid content of the volatile fraction (Fraction 2) was the lowest at 0.1 g / L as shown in Table 5 below.

sample Solids content, g / L Fraction 1 5.5 Fraction 2 0.1 Fraction 3 12.4 Fraction 4 3.6

3. Carbohydrate content analysis

Add 3 mL of 72% H ₂ SO ₄ to 10 mL of each fraction prepared in Example 1, add 79 mL of distilled water and 5 mL of inositol solution (20 mg / mL), and hydrolyze in an autoclave at 121 ± 3 ° C. for 1 hour . After being hydrolyzed, it was allowed to stand for 20 minutes, and the carbohydrate content was measured using gas chromatography (GC). At this time, pretreatment of the hydrolyzate for the carbohydrate content analysis was performed according to ASTM E1821-96, and GC analysis conditions were as follows.

Column: DB-225 (15 m x 250 m i.d., 0.25 m)

- Injection volume: 2 μl

- Injector temperature: 200 ℃

- Detector temperature: 250 ° C

Carrier gas: helium, 0.9 mL / min

- Split ratio = 1: 30

- Oven temperature: maintained at 190 캜 for 1 minute, then increased to 220 캜 at a rate of 10 캜 / min, maintained at 220 캜 for 10 minutes

As a result, the carbohydrate content of each fraction was confirmed as shown in Table 6 below. That is, five kinds of compounds were detected in the first liquid extract (Fraction 1), and the contents of gallic rosaceous and glucose were 16.2 g and 12.5 g, respectively, and arabinose had the lowest content of 0.9 g. The total carbohydrate content of the first liquid extract (Fraction 1) was 34.9%. The volatile fraction (Fraction 2) was collected from volatile components obtained from the process of concentrating the liquid phase of the oak hydrothermal extract, which had been subjected to the crushing treatment, and had no carbohydrate content.

The nonvolatile fraction (Fraction 3) was a non-volatile substance obtained by concentrating the first liquid extract (Fraction 1). Five kinds of compounds were detected, and the content of the germlose and glucose was 31.7 g and 23.5 g, 2.2%, respectively. The total carbohydrate content of the non-volatile fraction (Fraction 3) was 68.0%, indicating a carbohydrate content of about twice that of the non-volatile fraction (Fraction 3). The second liquid extract (Fraction 4) contained 1.4 g, 0.3 g of arabinose and jasilose, and the total carbohydrate content was 1.7 g. Diabetes mellitus was found to be a candidate for antidiabetic active ingredient in the volatile fraction (Fraction 2) in which the concentration of glucose in the blood was increased and the amount of carbohydrate was not detected.

sample Carbohydrate composition, g / 100 g Arabinose Jasil Ross Mannos Galactose Glucose Sum Fraction 1 0.9 16.2 2.6 2.7 12.5 34.9 Fraction 2 - - - - - - Fraction 3 2.2 31.7 5.3 5.3 23.5 68.0 Fraction 4 1.4  0.3 - - -  1.7

4. Analysis of mineral content

2 g of each fraction prepared in Example 1 was precisely weighed into a crucible and fully carbonized in an electric furnace at 600 占 25 占 폚. The crucible was allowed to dissipate on the asbestos plate for 2 minutes and then cooled in a desiccator. After adding a certain amount of 5% H ₂ O ₂ and repeatedly heating and cooling several times, the inorganic content was measured, and then the content was determined as W / S × 100, where W is the weight of the sample after carbonization (g), S is the weight of the sample (g)].

As a result, the inorganic content of the first liquid extract (Fraction 1) was 0.018%, and the volatile fraction (Fraction 2) was the volatile fraction and the inorganic content was the lowest as 0.005% as shown in Table 7 below. The mineral content of the non-volatile fraction (Fraction 3) was the highest at 0.020% and the mineral content of the second liquid extract (Fraction 4) was 0.013%.

sample Mineral content,% Fraction 1 0.018 Fraction 2 0.005 Fraction 3 0.020 Fraction 4 0.013

5. Analysis of polyphenol content

30 μl of each fraction prepared in Example 1, 3 ml of distilled water and 100 μl of Folin-ciocalteu reagent were added and reacted for 5 minutes. After 5 minutes, 300 μl of 20% Na ₂ CO ₃ was added to each test tube, Lt; / RTI > for 1 minute at room temperature. Absorbance was measured at 765 nm using a UV-spectroscopic analyzer, and the total polyphenol content was calculated by the following equation.

&Quot; (2) "

Total polyphenol content (%) = (sample absorbance + 0.0348) / 0.008

As a result, the non-volatile fraction (Fraction 3) showed the highest polyphenol content of 2989.8 mg / L and the second liquid extract (Fraction 4) had the polyphenol content of 1844.8 mg / L Respectively. The volatile fraction (Fraction 2) showed the lowest polyphenol content of 91.4 mg / L. Since polyphenol is a kind of antioxidant and antioxidant is known to have an effect on diabetes, it is considered that the fraction having a high total polyphenol content is a substance exhibiting an antidiabetic effect, and a nonvolatile fraction (Fraction 3) .

sample Total polyphenol content, mg / L Fraction 1 1271.8 Fraction 2   91.4 Fraction 3 2989.8 Fraction 4 1844.8

6. Analysis of organic acid content

1 mL of each fraction prepared in Example 1 was mixed with 3 mL of distilled water and 1 mL of Folin-Ciocalteu reagent 2N, reacted at room temperature for 5 minutes, and then 1 mL of 20% Na ₂ CO ₃ was added thereto, followed by reaction at room temperature for 1 hour . The reactants were analyzed for their organic acid content by measuring the absorbance at 640 nm using a UV spectrometer. The analyzed organic acids are vanillic acid, ferulic acid, and cinnamic acid. Vanillic acid can be obtained from vanillin which is oxidized or has a corrosive melting action. It is a substance of white and odorless bed, and ferulic acid is abundant in plant cell walls It contains an antioxidant activity that neutralizes free radicals and an effect of lowering blood sugar and cholesterol. Shinnam acid is one of typical aromatic unsaturated carboxylic acids. It has a phenyl group and a carboxy group on the opposite side to a double bond. It is a substance with a cis-isomer.

As a result, the content of the first liquid extract (Fraction 1) was 1.8 g / L of vanillic acid, 1.5 g / L of ferulic acid and 1.8 g / L of cinnamic acid and the volatile fraction (Fraction 2) 0.1 g / L of vanillic acid, 0.1 g / L of ferulic acid, and 0.1 g / L of cinnamic acid. The nonvolatile fraction (Fraction 3) showed the highest contents when compared with other fractions with vanillic acid 2.9 g / L, ferulic acid 2.5 g / L, and Shinnam acid 3.0 g / L. The second liquid extract (Fraction 4) contained 2.1 g / L of vanillic acid, 1.7 g / L of ferulic acid and 2.1 g / L of cinnamic acid. The fraction with the highest content of ferulic acid, which is antioxidant and hypoglycemic, was a nonvolatile fraction (Fraction 3).

sample Organic acid, g / L Vanillin acid Ferulic acid Shinnam-san Fraction 1 1.8 1.5 1.8 Fraction 2 0.1 0.1 0.1 Fraction 3 2.9 2.5 3.0 Fraction 4 2.1 1.7 2.1

≪ Example 3 > An antidiabetic test of an effective extract using an animal model

1. Inhibition of α-glucosidase activity

5 μl of each fraction prepared in Example 1 and 50 μl of α-glucosidase were mixed and allowed to react for 5 minutes, and the primary absorbance was measured at 405 nm by ELISA (Anthos ^® , Austria). Subsequently, 50 μl of substrate ( p -nitrophenyl-α-D-glucoside) was mixed and allowed to react for 5 minutes. Secondary absorbance was measured at 405 nm by ELISA (Anthos ^® , Austria).

As a result, as shown in the following Table 10, the volatile fraction (Fraction 2) showed a stronger inhibitory effect on alpha -glucosidase activity than the first liquid extract (Fraction 1). The non-volatile fraction (Fraction 3) showed about three times stronger α-glucosidase activity inhibitory effect than the second liquid extract (Fraction 4). At this time, when 10 μl of 5 mg / ml acavose was used as a positive control, the inhibition value was shown.

sample Akaboz Concentration (μl or mg / ml) 5 10 20 30 Fraction 1 23.45 + 0.04 24.2 ± 0.03 48.0 ± 0.09 78.7 ± 0.17 94.1 ± 0.00 Fraction 2 15.80 ± 0.03 28.2 ± 0.06 94.5 + 0.04 96.9 ± 0.03 97.4 ± 0.01 Fraction 3 15.76 + 0.02 33.1 + - 0.12 41.9 ± 0.05 62.3 + - 0.14 75.2 ± 0.07 Fraction 4 16.52 + 0.03 5.6 ± 0.02 9.1 ± 0.01 16.5 + 0.02 23.7 ± 0.01

2. Experiments on Type 1 diabetes using SD rats

Streptozotoxin was dissolved in 0.1 M citric acid buffer (pH 4.6) at 60 mg / kg in an experimental animal that was fasted for 12 hours on the day before, and male Sprague-Dawley (SD) Respectively. Forty-eight hours after the administration of streptozotocin, blood samples were collected from the tail vein, and blood glucose levels were determined to be more than 350 mg / dL.

To the prepared diabetic rats, the first liquid extract (Fraction 1) and the volatile fraction (Fraction 2) prepared in Example 1 were added to 0.1 ml per rat and the non-volatile fraction (Fraction 3) and the second liquid extract (Fraction 4) Each was administered orally at 200 mg / kg per rat, and metformin 500 mg / kg was orally administered as a control. Blood samples were collected from the tail vein at 30, 60, 120, and 180 minutes intervals, and blood sugar changes (mg / dl) were measured using a glucose meter.

As a result, as shown in the following Table 11, all the fractions showed reduced blood glucose levels as compared with the control group.

Sample 0 min 30 min 60 min 120 min 180 min Control 0 243 322 217 139 Fraction 1 0 28 245 165 64 Fraction 2 0 175 269 200 98 Fraction 3 0 138 172 171 94 Fraction 4 0 104 138 91 44 Metformin 0 147 138 40 -17

Example 4: Production of oak-fraction complex extract

(Fraction 1), a volatile fraction (Fraction 2), a nonvolatile fraction (Fraction 3), and a second liquid extract (Fraction 4), which are fractions prepared in Example 1, at various ratios And then each experiment was carried out. The related experiment was carried out in the same manner as in the previous embodiment.

1. Analysis of Total Polyphenol Contents by Mixing of Extracts of Oak Fraction Compositions

The total polyphenol content of each fraction was analyzed. As shown in Table 12, the volatile fraction (Fraction 2) having the lowest polyphenol content was obtained by mixing the non-volatile fraction (Fraction 3) .

Samples (mixing ratio, weight ratio) Total polyphenol content, mg / L Fraction 1: Fraction 3 (1: 1) 3127.8 Fraction 2: Fraction 3 (1: 1) 3011.8 Fraction 2: Fraction 4 (1: 1) 1899.2 Fraction 3: Fraction 4 (1: 1) 3254.3

2. Inhibitory effect of α-glucosidase activity on the mixture of oak extracts

As a result of examining the inhibition rate of? -Glucosidase activity according to the mixing of each fraction, the highest inhibitory rate of? -Glucosidase activity in a mixture of volatile fraction (Fraction 2) and non-volatile fraction (Fraction 3) Respectively.

Samples (mixing ratio, weight ratio) Concentration (μl or mg / ml) 5 10 20 30 Fraction 1: Fraction 3 (1: 1) 26.0 43.0 60.1 84.1 Fraction 2: Fraction 3 (1: 1) 31.0 70.2 78.2 89.3 Fraction 2: Fraction 4 (1: 1) 14.2 45.8 53.3 58.6 Fraction 3: Fraction 4 (1: 1) 18.1 23.3 40.2 48.3

As a result of examining the inhibition rate of α-glucosidase activity according to the mixing ratio of the volatile fraction (Fraction 2) and the non-volatile fraction (Fraction 3) showing the antidiabetic effect, the volatile fraction (Fraction 2 ) And non-volatile fraction (Fraction 3) were mixed at a weight ratio of 7: 3, the highest α-glucosidase activity was inhibited.

The sample (Fraction 2: Fraction 3, weight ratio) Concentration (μl or mg / ml) 5 10 20 30 1: 9 21.8 49.4 64.4 84.1 3: 7 23.6 51.4 68.7 87.8 7: 3 31.8 71.1 83.1 94.0 9: 1 28.8 62.2 79.4 92.0

Example 5: Toxicity test

The safety of the combined extract of oak fraction was evaluated by mixing the volatile fraction (Fraction 2) and the nonvolatile fraction (Fraction 3), which showed the most excellent antidiabetic activity, at a weight ratio of 7: 3.

First, the ICR mice (8-9 weeks old, 25 ± 3 g) were fasted for 10 hours, and then each of the females and males was orally administered once with the combination extract of the oak fraction at a concentration of 500, 1000 and 1500 mg / And general symptoms and body weight were measured for 14 days. Blood and pathological examination was performed by autopsy.

As a result, no clinical symptoms or dead animals were found in all animals, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings.

Hereinafter, various prescription examples including an extract of a combination of a volatile fraction (Fraction 2) and a non-volatile fraction (Fraction 3) at a weight ratio of 7: 3 as an active ingredient will be described. However, But rather just to explain in detail.

≪ Prescription Example 1 > Prescription Example of Pharmaceutical Composition

≪ Prescription Example 1-1 > Preparation of tablets

The extract was prepared by mixing 50 mg of the complex extract of oak fractions, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate, and a remaining amount of purified water, followed by tableting according to a conventional preparation method.

≪ Prescription Example 1-2 > Preparation of ointment preparation

After mixing 30 mg of the complex extract of oak fractions, 250 mg of PEG-4000, 650 mg of PEG-400, 10 mg of white petrolatum, 1.44 mg of methyl p-hydroxybenzoate, 0.18 mg of propyl p-hydroxybenzoate and the remaining amount of purified water, .

≪ Prescription Example 1-3 >

(70%), sucralose (30 mg), acesulfame potassium (60 mg), propylene glycol (1,000 mg), polyoxy 40 hydrogenated castor oil (500 mg), potassium sorbate (130 mg), sodium chloride (25 mg) 58 mg of citric acid, 26 mg of citrifluoroethane, and the remaining amount of purified water were mixed, and 100 ml of a liquid agent was prepared according to the usual method for producing a liquid agent.

≪ Prescription Example 2 > Prescription example of functional food

≪ Prescription Example 2-1 > Preparation of health food

(Vitamin E acetate, 70 mg of vitamin E, 1.0 mg of vitamin E, 0.13 mg of vitamin B 1, 0.15 mg of vitamin B 2, 0.5 mg of vitamin B 6, 0.2 mg of vitamin B 12, 10 mg of vitamin C, 10 mg of biotin, 1.7 mg of nicotinic acid amide, 50 엽 of folic acid, and 0.5 mg of calcium pantothenate), an appropriate amount of inorganic compound (ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium phosphate monobasic 15 mg, 55 mg of calcium, 90 mg of potassium citrate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride) were mixed to prepare a granule, and a health food was prepared according to a conventional method.

≪ Prescription Example 2-2 > Preparation of health drink

100 mg of citric acid, 100 g of oligosaccharide, 2 g of a plum concentrate, 1 g of taurine and purified water were added to make a total of 900 ml, and the above components were mixed according to a conventional health drink manufacturing method, After stirring for 1 hour at 85 ° C, the solution was filtered and sterilized in a sterilized 2 L container, and then refrigerated.

Claims (4)

70 to 90% by weight of a volatile fraction obtained from oak, and 10 to 30% by weight of a nonvolatile fraction obtained from oak,
The volatile fraction and the non-volatile fraction
Treating oak with high temperature and high pressure;
Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water;
Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And
And concentrating the liquid extract to obtain a volatile fraction and a non-volatile fraction, respectively. The method of claim 1, wherein the volatile fraction and the non-
The oak at a temperature of 190 to 250 ℃ at a pressure of 20 to 30 kgf / cm ² steps of high temperature and pressure for 2 to 30 minutes;
Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and subjecting it to hydrothermal extraction at a temperature of 55 to 65 ° C for 1 to 6 hours;
Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And
Wherein the liquid extract is obtained by concentrating the liquid extract at 75 to 85 캜 under vacuum for 1 to 3 hours to produce a volatile fraction and a non-volatile fraction, respectively. A pharmaceutical composition for the treatment or prevention of diabetes comprising as an active ingredient an extract of oak fraction comprising 70 to 90% by weight of a volatile fraction obtained from an oak and 10 to 30% by weight of a non-volatile fraction obtained from an oak,
The volatile fraction and the non-volatile fraction
Treating oak with high temperature and high pressure;
Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water;
Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And
And concentrating the liquid extract to obtain a volatile fraction and a non-volatile fraction, respectively, to obtain a pharmaceutical composition for treating or preventing diabetes. A functional food for diabetes mellitus improvement comprising as an active ingredient an oak fraction combination extract comprising 70 to 90% by weight of a volatile fraction obtained from an oak and 10 to 30% by weight of a non-volatile fraction obtained from an oak,
The volatile fraction and the non-volatile fraction
Treating oak with high temperature and high pressure;
Adding water to the oak which has been subjected to the high-temperature and high-pressure treatment and extracting it by hot water;
Removing the residue from the extract obtained by the hot water extraction to prepare a liquid extract; And
And concentrating the liquid extract to obtain a volatile fraction and a non-volatile fraction, respectively, to obtain a functional food for diabetes mellitus improvement.

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