JP2016183136A - Antioxidant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To aim at the improvement of the antioxidative effect in Black Soybean testa extract.SOLUTION: We have completed antioxidant, which is a present invention, from the finding that by a heat treatment of Black Soybean testa or Black Soybean testa extract in an acidic alcohol under the presence of divalent iron ion used as edibles, a product with high anti-oxidative effect can be obtained. Thus, the invention can suppress oxidative stress caused by aging, stress, biased eating habits, or the like.SELECTED DRAWING: None

Description

The present invention relates to the enhancement of the antioxidant effect of antioxidants taken orally, particularly black soybean seed coat extract.

In living organisms that breathe with oxygen, it is said that 1-2% of the breathed oxygen becomes active oxygen. Not only that, it is also subjected to oxidative stress due to active oxygen due to air pollution and external stimuli such as ultraviolet rays. Active oxygen produced in the body reacts with unsaturated fatty acids to produce lipid peroxides. From the aging of the skin to serious diseases such as Alzheimer's disease and cancer, the influence of active oxygen is extremely large, and one theory is that active oxygen is involved in 85% of all diseases. Normally, the generated active oxygen is removed by the action of antioxidant enzymes in the body. However, if the activity of the antioxidant enzymes is weakened or excessively generated, the active oxygen that cannot be erased is considered a harmful substance. Damages various parts of the body. In order to suppress such damage caused by active oxygen and various diseases, it is effective to suppress the generation of active oxygen itself.

Therefore, by using a monovalent or divalent metal ion in combination with a plant extract having antioxidant power, a method for enhancing the antioxidant action and enhancing the stability (Patent Document 1), catechins and procyanidins A method (Patent Document 2) is known that provides an antioxidant having an excellent antioxidant action by heat treatment at a high temperature. Also disclosed is a method for improving the antioxidant effect by hydrolyzing black soybean extract (Patent Document 3). In this document, not only enzymes but also hydrolysis methods using acids or alkalis are shown.

JP 2001-139484 A JP2007-254508 JP 2006-290742 A

Polyphenols such as catechins and procyanidins, which are plant components, are known to capture active oxygen by OH groups in the chemical structure and have a strong antioxidant action. Plants such as black soybeans contain a large amount of procyanidins, and seed coat extracts are known to have a strong antioxidant effect. There are many health foods and pharmaceuticals that utilize the antioxidant action of polyphenols, and they are widely used around the world. However, polyphenols have a low bioabsorption rate and a short residence time. Therefore, in order to obtain an effective antioxidant effect, frequent and large-scale intake is required. Therefore, in order to solve these drawbacks, it is desired to develop a product having a higher antioxidant property.

As a result of intensive studies, the inventors of the present invention have high antioxidant power in products obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol in the presence of divalent iron ions. I found out.

(I) Antioxidant I-1. An antioxidant comprising a product obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol in the presence of divalent iron ions as an active ingredient.
I-2. The antioxidant described in 1-1, which has an oral dosage form.
I-3. The anti-antibody described in either I-1 or I-2 having a pharmaceutical form selected from tablets, pills, capsules (hard capsules, soft capsules), powders, granules, troches, liquids, and jellys Oxidant.
I-4. The antioxidant described in any one of I-1 to I-3 used in combination with food and drink, preferably water or fresh water.

(II) Additive for oral composition having antioxidant action II-1. An additive for an oral composition comprising as an active ingredient a product obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol in the presence of divalent iron ions.
II-2. The additive for oral compositions described in II-1 which is an additive for food and drink.
II-3. Oral as described in either II-1 or II-2 having a pharmaceutical form selected from tablets, pills, capsules (hard capsules, soft capsules), powders, granules, troches, liquids, and jellys Additive for composition.

The antioxidant of this invention can obtain the product which has high antioxidant power by heat-processing black soybean seed coat or black soybean seed coat extract in acidic alcohol in presence of a bivalent iron ion. Moreover, the additive for oral compositions which has an antioxidant action of this invention can be used in order to strengthen an antioxidant action with respect to an oral composition. That is, the additive of the present invention can be used for the preparation of an oral composition having an antioxidant action, and thus suppresses or improves oxidative stress caused by aging, stress, unbalanced diet, etc. An oral composition (oral drug, oral quasi-drug, food or drink) having an antioxidative effect with excellent effects can be prepared.

(I) Antioxidant The antioxidant of the present invention contains, as an active ingredient, a product obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol with addition of divalent iron ions. It is characterized by that.

The black soybean used in the present invention is classified into the leguminous soybean genus (Glycine max (L.) Merrill) in terms of plant taxonomy, but the appearance of the seed is black and contains anthocyanin pigments. It is distinct from soybeans in that it has strong antioxidant properties. There are varieties of black soybeans, such as Tamba Black, Purple Riding Hood, Meiko Kurokuro, Tokachikuro, Iwakuro, Tamadakuro, Tamba Black, Shinano Black, etc. Also good.

In the present invention, black soybean can be used as a raw material for processing as it is. Further, it may be powdered by crushing, grinding or the like. In preparing an extract from black soybean, it is most preferable to use only seed coat from the viewpoint of having a strong antioxidant effect. The seed coat can be washed with water in advance to remove foreign substances (raw or dried product), if necessary (crushed product, crushed product, and powdered product). Good.

(A) When using black soybean seed coat extract The method for extracting the active ingredient from the black soybean seed coat is not particularly limited, but it can be carried out by contacting with an extraction solvent. Specifically, the active ingredient can be extracted by a method such as immersion in a solvent and stirring.
There are no particular limitations on the temperature conditions used for extraction, and any of low temperature, normal temperature, and heating conditions (including high temperatures) may be used, but heating conditions (including high temperatures) are preferred. More specifically, when extracting with the below-mentioned hydrous lower alcohol, it is 30 degreeC or more, Preferably it is the range of 40 to 60 degreeC. The extraction time under such temperature conditions is not limited, but it may be 60 minutes or longer, preferably 90 to 120 minutes. When extracting with water or acidic aqueous solution, temperature is 40 degreeC or more, Preferably it is the range of 60-80 degreeC, and extraction is performed for 10 minutes or more on this temperature condition, Preferably it is about 20 minutes-120 minutes.

The water-soluble solvent used for extraction is not particularly limited, and examples thereof include water, lower alcohols, and mixtures thereof. Examples of lower alcohols include C1-C4 lower alcohols such as methanol, ethanol, propanol, isopropyl alcohol, and butanol. Preferred examples of the lower alcohol include ethanol. The water-soluble solvent is preferably a water-containing lower alcohol (particularly water-containing ethanol) or water. In addition, when using a hydrous lower alcohol as a solvent, it is preferable that the amount of lower alcohol contained in it is 80 volume% or less.

The water-soluble solvent used for extraction may be adjusted to be acidic. Although not particularly limited, the pH range of the water-soluble solvent is preferably about pH 1 to 5. In order to adjust the pH of the water-soluble solvent to be in such a range, an appropriate acidic substance such as an inorganic acid or an organic acid can be usually used.

Specific examples of acidic substances include inorganic acids such as hydrochloric acid, sulfuric acid, and nitric acid; and organic acids such as methanesulfonic acid, benzenesulfonic acid, (above, sulfonic acid), formic acid, acetic acid, and (above, carboxylic acid). Can be mentioned. An acid having a sulfo group is preferable, and sulfuric acid is particularly preferable. The normality of the acid in the water-soluble solvent is not particularly limited as long as the water-soluble solvent is in the above pH range, but is preferably in the range of 0.01 to 0.5 N, more preferably 0.03. It is in the range of ~ 0.5N.

As long as the effects of the present invention can be achieved, the obtained black soybean seed coat extract is further subjected to purification treatment such as filtration, coprecipitation or centrifugation to remove solids, extraction treatment, adsorption treatment, etc. May be.

The obtained black soybean seed coat extract is neutralized if necessary, further diluted or concentrated to an appropriate concentration, insolubles are removed by filtration, etc., and freeze drying, spray drying, low temperature drying, etc. You may powder according to a method.

In preparing a heat-treated product containing divalent iron ions from the black soybean seed coat extract prepared as described above, the black soybean seed coat extract is dispersed or dissolved in acidic alcohol, and divalent iron is added. Heat in the presence of ions. Although there is no restriction | limiting in particular in the dose of acidic alcohol in that case, Preferably it is about 10 to 1000 times amount with respect to the weight of a black soybean seed coat extract.

The solvent used is adjusted to be acidic. Specific examples of acids include inorganic acids such as hydrochloric acid, sulfuric acid, and nitric acid; and organic acids such as methanesulfonic acid, benzenesulfonic acid (above, sulfonic acid), formic acid, and acetic acid (above, carboxylic acid). Can do. Of these, hydrochloric acid is preferred. Although the concentration of hydrochloric acid in the solvent is not particularly limited, it is preferably 0.1% or more and 10% or less in view of the subsequent neutralization.

Examples of the alcohol used include lower alcohols and polyhydric alcohols, and the above-mentioned acids can be mixed and used. Specifically, lower alcohols include methanol, ethanol, propanol and the like, and polyhydric alcohols include ethylene glycol, propylene glycol, glycerin and the like.
In the present invention, among them, a lower alcohol is suitably used, preferably methanol or ethanol, and more preferably ethanol in view of use in foods. In the case where a lower alcohol is used as a solvent, the concentration is not particularly limited, but is 10% or more, preferably 50% or more.

Examples of the divalent iron ion to be added include iron (II) sulfate and iron (II) ammonium sulfate, and iron (II) sulfate is preferable in view of use in foods. The concentration at the time of use is not particularly limited, but in the case of oral administration, it is preferably about 0.01 to 1% in order to prevent excessive intake of iron.

The heating temperature is in the range of 50 to 100 ° C., preferably about 80 to 100 ° C. under normal pressure. The heating time is not particularly limited, but if the heating time is too long, it may cause a decrease in the antioxidant effect, and therefore it is preferably 30 minutes or longer and 6 hours or shorter.

The product obtained by the above heat treatment is diluted or concentrated as necessary to an appropriate concentration, and insoluble matter is removed by filtration or the like, followed by lyophilization, spray drying, low temperature drying, etc. May be used.

(B) When using black soybean seed coat In preparing a heat-treated product containing divalent iron ions from black soybean seed coat, black soybean seed coat is dispersed in acidic alcohol and heated in the presence of divalent iron ions. . Although there is no restriction | limiting in particular in the dose of acidic alcohol in that case, Preferably it is about 10 to 1000 times amount with respect to the weight of a black soybean seed coat.

The type of black soybean used, the method for obtaining black soybean seed coat, the method for preparing acidic alcohol, the type of iron ion, the heating conditions (temperature and time), and the method for preparing the heat-treated product are as described above (А). Yes, it can be used in this column (B).

The heat-treated product prepared in the above (A) or (B) can be used as the antioxidant of the present invention regardless of its form as long as it is an oral administration form. In addition, as long as it has an oral administration (oral intake) form, it is classified according to its use (pharmaceuticals, quasi drugs, foods and drinks [including health functional foods and supplements such as foods for specified health use and nutritional functional foods] ) Is not particularly limited.

Oral dosage forms include liquids (including extract forms and syrups) or jelly forms; powders or fine powders prepared by the above heat treatment into powders or granules by conventional methods. Examples include granules or granules; capsules filled with liquids, powders or granules (hard capsules, soft capsules); or tablets or powders that are further compressed into tablets. Yes (solid formulation)

The antioxidant of the present invention is adjusted to various dosage forms (oral dosage forms) by combining the black soybean seed coat extract with pharmaceutically or food-acceptable edible carriers, excipients and the like. You can also.

(II) Additive for oral composition having antioxidant action The additive for oral composition having antioxidant action of the present invention is an acidic alcohol obtained by adding divalent iron ions to black soybean seed coat or black soybean seed coat extract. The product obtained by heat-treating in is used as an active ingredient.

The additive of the present invention is targeted by a product obtained by heat-treating black soybean seed coat or black soybean seed coat extract, which is an active ingredient, in acidic alcohol in the presence of divalent iron ions. It is an additive used for imparting an antioxidant action to an oral composition. The additive of the present invention can also be used to further enhance the action of an oral composition having an antioxidant action.

Here, the oral composition targeted by the present invention includes a composition that is orally administered to a person or a composition that is taken by a person, specifically an oral pharmaceutical composition, an oral quasi-drug composition. And food compositions (including beverages, hereinafter the same). A food composition is preferred.

The type of black soybean used as a raw material for the additive of the present invention, the method for obtaining black soybean seed coat, the method for preparing black soybean seed coat extract, the method for preparing acidic alcohol, the type of iron ion, the heating conditions (temperature and time), The method for preparing the heat-treated product of black soybean seed coat extract is as described in (I) above, and can be incorporated in this section (II).

The additive of the present invention may be a product itself obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol in the presence of divalent iron ions, or pharmaceutically. Further, it may be prepared by combining conventionally known edible carriers, excipients and the like that are acceptable as food. The additive of the present invention is used to prepare a composition (oral composition) having an antioxidative action by being added to and blended with the above oral medicine, oral quasi-drug, and / or food and drink. Therefore, as long as the form is not particularly limited, even if it has a liquid form (including extract form and syrup) and a jelly form, the liquid form is formulated into a powder or granule by a conventional method. Powders, fine granules, granules; capsules filled with liquids, powders or granules (hard capsules, soft capsules), or powders or granules further compressed into tablets It may be (solid preparation).

The amount of the additive of the present invention used by being added to oral pharmaceuticals, quasi-drugs, and / or foods and drinks is an oral composition having an antioxidant action prepared by adding the additive of the present invention. Daily administration (ingestion) of the product can be arbitrarily determined.

The additive of the present invention is used as one of raw materials together with other raw materials in the step of preparing the above oral composition (oral medicine, oral quasi-drug, and / or food or drink), or orally. When the composition is taken (administered or ingested), it can be used in the oral composition after use.

Hereinafter, the present invention will be described in more detail with reference to experimental examples. The antioxidant action is indicated by POV, but the scope of the present invention is not limited to this.

Experimental example Evaluation of antioxidant effect of heat-treated product containing divalent iron ions of black soybean seed coat extract The outline of a specific experiment is shown below.

(1) Preparation of sample (a) Preparation of black soybean seed coat extract The black soybean seed coat extract was prepared by the following method.
(1) 500 g of black soybean seed coat (raw) was immersed in 15 L of 40% ethanol aqueous solution as an extraction solvent, and extracted with stirring for 90 minutes while heating to 40 ° C.
(2) After extraction, the black soybean seed coat and liquid were separated into solid and liquid, and the liquid was recovered.
(3) The collected liquid was centrifuged to remove suspended solids that could not be removed by solid-liquid separation.
(4) The resulting extract was evaporated by heating under reduced pressure and concentrated.
(5) Dextrin was added as an excipient to the resulting concentrated solution.
(6) This was heat-sterilized and then dried to a powder (total 119.8 g, containing black soybean seed coat extract 31.9 g).

(B) Preparation of heat-treated product containing divalent iron ions of black soybean seed coat extract 1000 mg of the above extract was dissolved in 1 L of 0.5% hydrochloric acid / ethanol, and 2% as divalent iron ions was added to this solution. 30 ml of an aqueous solution of iron (II) sulfate or 2% aqueous solution of iron (II) ammonium sulfate hexahydrate was added. After thoroughly stirring and mixing, the mixture was heated at 100 ° C. for 2 hours. Furthermore, what was powdered by concentrating and freeze-drying was used as a sample (Examples 1 and 2).
Moreover, as a comparative control, the heat-treated product of black soybean seed coat extract is Comparative Example 1, the mixture of black soybean seed coat extract and iron (II) sulfate is Comparative Example 2, the untreated black soybean seed coat extract is Comparative Example 3, Comparative Example 4 was obtained by heating iron (II) sulfate in acidic alcohol.

(2) Experimental method In this test, the antioxidant effect was evaluated by the lipid peroxide suppression method by POV measurement of linoleic acid.

To 20 ml of ethanol containing 500 mg of linoleic acid, 26 ml of 0.1 M phosphate buffer adjusted to pH 7.0 in advance was added. 2 ml of what dissolved each sample of Examples 1-2 and Comparative Examples 1-4 in the 0.5% hydrochloric acid methanol solution was added here. 2 ml of 0.05% magnesium chloride aqueous solution was added and incubated in the dark at 45 ° C. for 7 days. To the sample after completion of the incubation, 2 ml of 6N hydrochloric acid was added and extracted twice with 10 ml of chloroform to obtain a chloroform layer. Acetic acid (5 ml) and saturated potassium iodide aqueous solution (1 ml) were added to 10 ml of the chloroform layer thus obtained and left in the dark for 5 minutes. 1% starch solution was added and titrated with 0.01M sodium thiosulfate solution, and the point when the color disappeared was taken as the end point. A sample to which a test sample was not added was prepared as an empty test group, and POV was analyzed without incubation. POV was calculated by the following formula. The results are shown in FIG.
POV (meq / kg) = (A−B) × F × 10 / oil sample amount A: titration (ml)
B: Titration volume of empty test area (ml)
F: Sodium thiosulfate titer

(3) Experimental Results FIG. 1 shows the POV analysis results of Examples 1-2 and Comparative Examples 1-4.

Regarding the antioxidant property of the antioxidant of the present invention, the amount of lipid peroxide produced by the oxidation of linoleic acid was measured by the POV method. The POV value indicates that the smaller the number, the more the oxidation is suppressed.
As shown in FIG. 1, it can be seen that Examples 1 and 2 are clearly superior in lipid peroxide generation suppression effect by oxidation of linoleic acid as compared with Comparative Example 1. That is, a product having high antioxidant power can be obtained by heat-treating black soybean seed coat extract in the presence of divalent iron ions.

Antioxidant effect of heat-treated product containing divalent iron ions of black soybean seed coat extract

Claims (2)

An antioxidant comprising a product obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol in the presence of divalent iron ions as an active ingredient. An additive for an oral composition comprising as an active ingredient a product obtained by heat-treating black soybean seed coat or black soybean seed coat extract in acidic alcohol in the presence of divalent iron ions.