JP2006057012A - Antioxidative composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an antioxidative composition having antioxidative effect which is useable for a wide range of foods and beverages and medicines, and foods and beverages, quasi-drugs and medicine each containing the composition in order to solve problems in which it is required to take an antioxidation substance in the human body in order to prevent attack of cell membrane by active oxygen and eliminate the active oxygen, because about 2%-4% of oxygen molecules taken by human respiration become active oxygens having unstable unpaired electrons and those active oxygens attack the cell membranes in order to obtain stability themselves and further when SOD is orally taken in order to supplement insufficient SOD in the human body, it is difficult to rapidly absorb SOD and orally taken SOD is decomposed by gastric acid and activity of SOD is lost and it is required as effective means to take antioxidation substance exhibiting activity similar to that of SOD. <P>SOLUTION: The antioxidative composition comprises Amla (Emblica officinale or Phyllanthus embilica) or its extract. In the antioxidative composition, the extract of Amla is obtained by extracting Amla fruit or juice with either one of water, a base, an acid, a hydrophilic solvent and acetone. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an antioxidant composition containing Amlar fruit, fruit juice, or an extract thereof, and a food, beverage, quasi-drug, and pharmaceutical product containing the same.

  Humans breathe constantly and take oxygen into their bodies. Approximately 2% to 4% of oxygen ingested by breathing is an active oxygen with unstable unpaired electrons (those with one or more unpaired electrons in the molecule are called free radicals, and oxygen free radicals are active oxygen. These active oxygens attack cell membranes for stability in an attempt to stabilize quickly. Thereafter, the unsaturated fatty acid in the cell membrane is oxidized, and the oxidized unsaturated fatty acid is converted into a lipid peroxyl radical. The lipid peroxyl radical again oxidizes unsaturated fatty acids and generates lipid peroxides while repeating oxidation. Although active oxygen also has a role of protecting itself from bacteria that have invaded the body, the problem is when a large amount of active oxygen is generated. Active oxygen is harmful because unsaturated fatty acids are oxidized by active oxygen in the body to become “lipid peroxide”. The human body is composed of approximately 60 trillion cells, and the outermost cell membrane is made of “unsaturated fatty acids”. When the unsaturated fatty acid is oxidized to become lipid peroxide and the cell membrane becomes tattered, even proteins in the cell membrane are involved and oxidized to form a more harmful secondary oxide. As a result, various diseases such as aging, stroke, autoimmune disease, cancer, cataract, fatty liver, myocardial infarction, ischemic heart disease, Alzheimer syndrome, diabetes, diabetic nephropathy, arteriosclerosis, allergy, hay fever and Aging such as skin wrinkle formation and skin elasticity decrease, inflammation, and skin pigmentation are caused. Fortunately, however, the human body is equipped with antioxidants, and it contains antioxidants such as SOD (superoxide dismutase), catalase and glutathione peroxidase in the body. Give me. However, this SOD also decreases with age. Moreover, due to many environmental pollutants such as recent agricultural chemicals, insecticides, food additives, nitrogen compounds, ultraviolet rays, radiation, stress, tobacco, environmental pollution only consumes precious SOD in our bodies. There is no environment where active oxygen is made.

  In order to eliminate active oxygen, it is only necessary to take an antioxidant, and if SOD in the body is insufficient, SOD may be supplemented, but it is difficult to absorb SOD quickly even if taken orally, In addition, it is decomposed into stomach acid and loses its activity. Therefore, taking an antioxidant substance exhibiting the same activity as SOD is an effective means.

  Examples of antioxidants include water-soluble antioxidant compounds such as L-ascorbic acid (vitamin C), reduced glutathione, uric acid, polyphenols, α-tocopherol (vitamin E), ubiquinone (coenzyme Q), carotenoids And the like, and synthetic compounds such as BHT (butylhydroxytoluene) and BHA (butylhydroxyanisole) are known. However, synthetic antioxidants BHT and BHA have problems such as suspected carcinogenicity. Considering the daily intake of antioxidants, foods are mainly used as raw materials in the production of antioxidants because of their economics and safety. For example, polyphenols such as catechins from tea leaves and procyanidins from red wine Kinds are manufactured. Antioxidants as other natural ingredients include extracts selected from plums, edibles, bengal karatachi, miracle berries, yaba santa, lemongrass (for example, see Patent Document 1), agaricus, mesimacob, and propolis ( For example, refer to Patent Document 2), spikelet extract (for example, refer to Patent Document 3), salamander aqueous solvent extract (for example, refer to Patent Document 4), cacao leaf, bark or root extract (for example, , See Patent Document 5), extract of bran, germ or flour with organic solvent (see, for example, Patent Document 6), vegetables that are said to be nutritionally superior, pumpkin, spinach, morroheia, tomato, What consists of 15 or more types of natural food materials, such as sweet corn, carrot, purple corn, is known (for example, refer patent document 7). On the other hand, it has been found that ingesting various and various antioxidants in small amounts is more effective in preventing various diseases than ingesting large amounts of a single substance. For that purpose, it is necessary to provide various antioxidants.

JP 2003-183120 A (page 1-2) JP 2002-235084 A (page 1-2) JP 2002-371092 A (page 1-2) JP 2002-104985 A (page 2-3) JP 2000-60485 A (page 1-2) JP-A-10-147779 (page 1-2) JP-A-10-75740 (page 2-5)

  An object of the present invention is to provide a composition having an antioxidant effect that can be used for a wide variety of foods and drinks and pharmaceuticals, and foods and drinks, quasi drugs and pharmaceuticals containing the same.

  In view of such circumstances, the present inventors have intensively studied to improve the problems of the prior art, and as a result, found that Amler has a strong antioxidant action and SOD-like activity, and completed the present invention. It was.

Antioxidant composition containing Amler fruit, fruit juice or extract thereof obtained in the present invention is measured for DPPH (diphenylpicrylhydrazyl) free radical scavenging ability using an electron spin resonance (ESR) apparatus. Results and SOD-like activity measurement results and streptozotocin (STZ) obtained by capturing reactive oxygen species generated by an appropriate method with a spin trap agent such as DMPO and quantifying the ESR signal specific to each reactive oxygen species Substance that reacts with thiobarbituric acid (TBA) with lipid peroxide increased in vivo by oxidation by administration of antioxidant composition to rats with diabetic nephropathy induced by administration and addition test to fish meat From the result of measurement with the amount of (thiobarbituric acid-reactive substance; TBARS), it was found that the antioxidant effect was high.
In particular, Amler is a natural plant that humans have been using for a long time in their daily diets, and unlike conventional drugs, it is safe with no side effects.
The present invention uses an antioxidant composition containing Amla fruit, fruit juice or an extract thereof for various foods and medicines, aging, stroke, autoimmune disease, cancer, cataract, fatty liver, myocardial infarction, Suppresses the progression of various diseases such as ischemic heart disease, Alzheimer's syndrome, diabetes, diabetic nephropathy, arteriosclerosis, allergy, hay fever and abnormal pigmentation such as stains and freckles, and the deterioration of food Can do.

  Amler used in the present invention is the scientific name: Emblica officinale or Philanthus embrica, which is a sub-tree of deciduous trees belonging to the genus Euphorbiaceae, from the Indian region to southern Malaysia and China. It is distributed over the country, and India is considered the origin. In addition, each region or language has its own unique name, such as citrus, oil sweets, Satsuma mushrooms, Emblic Mirobaran, Armala Key, Melaka tree, Malacca tree, Indian Gooseberry, Aronra, Amira, Amiraki, Amira Cattra, Nerikai, It is also called Neruri, Tasha, Kayuraka, Kemuraka, Nakhon Pong and others.

In the present invention, the site of Amler is not particularly limited, and fruits, fruit juices, leaves, stems, roots and the like can be used, but fruits and fruit juices are preferably used. The form is not particularly limited, and any of immature fruit, ripe fruit, dried fruit, fruit juice, fruit juice powder, and the like may be used.
In the case of fruit juice or fruit juice powder, it can be used as it is, but when using a product containing a water-insoluble component such as fresh fruit or dried fruit, it is effective that the water-insoluble component is removed by extraction. Is preferable.
At the time of extraction, when using fresh fruits, it is preferable to use seeds that have been crushed and homogenized with a mixer or the like in order to increase extraction efficiency, with or without the addition of water.
When using a dried fruit, it is preferable to grind | pulverize so that it may become a particle size of 40 mesh or less, in order to improve extraction efficiency.

The extraction method is not particularly limited, such as extraction solvent and extraction temperature, and water, base, acid, hydrophilic solvent, and acetone can be used as the extraction solvent. The hydrophilic solvent is preferably one or more selected from the group of lower alcohols such as methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, and butyl alcohol from the viewpoint of operability and extraction efficiency. Particularly preferred is water, base, or acid.
When using an acid or a base as the extraction solvent, it is preferable to neutralize the extract. The salt produced by the neutralization reaction can be removed by a known method such as dialysis or gel filtration. When water is used as the extraction solvent, the neutralization reaction as described above is not necessary, and it is not necessary to remove the generated salt. Therefore, it is more preferable to use water.
The acid used at this time is not particularly limited, and most of the acid can be used. However, it is preferable to use one kind or a combination of both selected from hydrochloric acid and sulfuric acid because of easy availability and operability. is there.
Further, the base is not particularly limited, and most of the bases can be used, but preferably one kind selected from sodium hydroxide and potassium hydroxide or a combination of both.
The concentration of the acid or base used for the extraction is not particularly limited before or after the extract is treated with the enzyme, and varies depending on the strength of the acid or base. From the viewpoint of efficiency, it is preferable to use a concentration of 0.01 to 0.5 mol.

The above fruit juice or extract can be used as it is, but it is preferable because the antioxidation effect is enhanced and the application range is widened by removing insoluble substances and solvent by filtration, centrifugation and fractional distillation.
After removing the insoluble substances and the solvent, the fruit juice or the extract is directly or concentrated and then distributed using an organic solvent to obtain each solvent-soluble fraction. These solvent-soluble fractions are preferred because the antioxidant effect is further enhanced. The organic solvent is selected from the group consisting of methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, butyl alcohol lower alcohol, ethyl acetate, butyl acetate, diethyl ether, methyl ether, methyl isobutyl ketone, hexane, acetone, or chloroform. One or more types can be used. In order to increase the purity of the soluble fraction, partitioning with other hydrophobic solvents can be combined. The concentration of these solvents is not particularly limited, but is preferably 10 to 100%, more preferably 60 to 100% as the final concentration in terms of yield and effects.
In order to further increase the purity, it is also possible to use a hydrophobic resin based on phenol, styrene, acrylic acid, epoxyamine, pyridine, methacryl or the like. In that case, as an eluent after resin adsorption, lower alcohols such as methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, and butyl alcohol, and acetone can be used alone or as an aqueous solution.
The extract and fraction can be used as they are, but if necessary, they can be used after being dried and powdered by means such as spray drying or freeze drying.

  The antioxidant effect can be confirmed by measuring free radical scavenging ability and lipid peroxide in blood. The measurement of free radical scavenging ability is carried out by EDP, chemical method such as cytochrome C reduction method, nitrous acid method, nitro blue tetrazolium salt reduction method (NBT reduction method), DPPH radical (diphenylpicrylhydrazyl radical). ) The inhibition rate, superoxide radical scavenging ability, SOD-like activity value, and hydroxy radical value can be confirmed by measurement. Although the effect is not particularly limited, usually, the SOD-like activity value is 1 to 6, the DPPH inhibition rate is 30 to 100%, preferably, the SOD-like activity value is 4 to 6, and the DPPH inhibition rate is 50 to 100. % Is good.

The antioxidant composition of the present invention can be applied to foods and drinks, pharmaceuticals, feeds and the like, and preferably foods and drinks, quasi drugs and pharmaceuticals that can be easily consumed by humans.
The food and drink in the present invention is not particularly limited as long as it is a form that can be taken orally, such as a solution, suspension, powder, or solid molded product. More specifically, instant noodles, retort foods, canned foods, microwave foods, instant soups and miso soups, freeze-dried foods, soft drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks Beverages such as powdered beverages, concentrated beverages, nutritional beverages, alcoholic beverages, bread, pasta, noodles, cake mixes, flour products such as fried flour and bread crumbs, rice cakes, caramel, chewing gum, chocolate, cookies, biscuits, cakes, Sweets such as pies, snacks, crackers, Japanese sweets, desserts, sauces, processed tomato seasonings, flavor seasonings, cooking mixes, sauces, dressings, soups, curry and stew seasonings, processing Fats and oils such as butter, margarine and mayonnaise, milk beverages, yogurts, lactic acid bacteria beverages, ice creams, chestnuts Agricultural processing of dairy products such as mussels, frozen foods, processed fishery products such as fish ham and sausages, marine products, processed livestock products such as livestock ham and sausages, canned agricultural products, jams and marmalades, pickles, boiled beans and cereals Products, nutritional foods, tablets, capsules and the like.

  The intake amount of the antioxidant composition of the present invention as a food or drink should be adjusted according to the disease state of the present invention, the body weight, age, constitution, physical condition, etc. of the sick person, but in general, the antioxidant composition per day The product can be appropriately selected in the range of 0.05 g to 20 g, preferably 0.1 g to 5 g. This can be taken once or several times a day depending on the state of the disease or the form of food.

In this invention, when processing into an antioxidant composition or food-drinks, various nutrient components can be strengthened.
Nutritional ingredients that can be strengthened include vitamins such as vitamin A, vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , vitamin B ₁₂ , vitamin C, vitamin D, vitamin E, niacin (nicotinic acid), pantothenic acid, folic acid, Essential amino acids such as lysine, threonine and tryptophan, minerals such as calcium, magnesium, iron, zinc and copper, and for example, α-linolenic acid, EPA, DHA, evening primrose oil, octacosanol, casein phosphopeptide (CPP), Casein calcium peptide (CCP), water-soluble dietary fiber, insoluble dietary fiber, substances that contribute to human health, such as oligosaccharides, and one or more useful substances approved as other foods and food additives Can be used.

The quasi drugs and pharmaceuticals in the present invention can be prepared as oral preparations or injections according to conventional methods using excipients and other additives suitable for oral or parenteral administration. Preferred is an oral preparation, and most preferred is an oral solid preparation that can be easily taken and is easy to store and carry.
As oral solid preparations, tablets, powders, fine granules, granules, capsules, pills, sustained-release preparations and the like are used. In such solid preparations, appropriate pharmacologically acceptable carriers, excipients (eg starch, lactose, sucrose, calcium carbonate, calcium phosphate etc.), binders (eg starch, gum arabic, carboxymethylcellulose, Mixed with hydroxypropylcellulose, crystalline cellulose, alginic acid, gelatin, polyvinylpyridone, etc.), lubricant (eg, stearic acid, magnesium stearate, calcium stearate, etc.), disintegrant (eg, carboxymethylcellulose, talc, etc.), etc. Tablets, powders, fine granules, granules, capsules, pills, sustained release agents, etc. can be prepared by the method. Oral liquid preparations include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, etc., and generally used inert diluents such as purified water and ethyl alcohol. Including. In addition to the inert diluent, the composition may contain adjuvants such as wetting agents and suspending agents, sweeteners, flavors, fragrances and preservatives.
Examples of injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of the aqueous solution and suspension diluent include distilled water for injection and physiological saline. Examples of diluents for water-insoluble solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethyl alcohol, polysorbate 80, and the like. Such compositions may further contain adjuvants such as preservatives, wetting agents, emulsifiers, dispersants, stabilizers (eg lactose), solubilizers (eg glutamic acid, aspartic acid). These are sterilized by, for example, filtration through a bacteria storage filter, blending with a bactericide or irradiation. These can also be used by producing a sterile solid composition and dissolving it in sterile water or a sterile solvent for injection before use.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, the following Example does not limit the scope of the present invention.

(Example 1) Preparation 1 of an antioxidant composition
Amla dried fruit was pulverized to 40 mesh or less, 2 L of distilled water was added to 80 g of the powder, and extracted at 55 ° C. for 3 hours. Then, it centrifuged (3000 rpm, 10 minutes), the supernatant was filtered, and the extract and the residue were isolate | separated. Distilled water (2 L) was added to the residue, and the mixture was repeatedly extracted under the same conditions. The extracts were combined and then lyophilized to obtain 35.0 g of the antioxidant composition A of the present invention. The yield was 43.8%.

(Comparative example 1) Preparation of various extracts 1 L of warm water of 70 ° C was added to 50 g of Kabuse Ise tea, and extracted by shaking for 5 minutes. Thereafter, the mixture was centrifuged, the supernatant was filtered, and the filtrate was concentrated under reduced pressure and lyophilized to obtain 18.4 g of a tea extract as a comparative sample. Further, 17.6 g of oolong tea extract was obtained from 50 g of oolong tea (made in Taiwan) by the same method. Furthermore, the grape seed extract was obtained from Sabinsa (made in India). Vitamin C was obtained from Wako Pure Chemicals as reagent grade.

(Test Example 1) Superoxide radical measurement by ESR Superoxide radical of SOD reagent (15000 Unit: Labotech) by spin trap method using ESR spectrometer (JES-RE1X (X-band): manufactured by JEOL Ltd.) (O ²⁻ ) was measured.
The SOD reagent was diluted with distilled water to prepare samples having respective concentrations of 0, 0.78125, 1.5625, 3.125, 6.25, and 12.5 U / mL. 50 μL of 2 mM hypoxanthine (manufactured by Labotech) adjusted with phosphate buffer (pH 7.8) in a test tube for ESR, 5.5 mM DTPA (Diethylene triamine acetate acid: Labtech) adjusted with phosphate buffer (pH 7.8) 35 μL, sample 50 μL, 0.092 μM DMPO (5,5 Dimethyl-1-pyrroline-1-oxide) 15 μL adjusted with distilled water and xanthine oxidase 50 μL adjusted with phosphate buffer (pH 7.8) were added to add 10 Mixed for minutes. Thereafter, the ESR signal was quantified under the following measurement conditions. Mn ²⁺ was used as an internal standard. The results are shown in Table 1.
ESR measurement conditions: magnetic field 336 mT, modulation frequency 100 KHz, modulation width 0.1 mT, response time 0.03 seconds, output 8 mW, sweep time 40 seconds

  As shown in Table 1, the IC50 showing the 50% SOD activity value of the SOD reagent was 0.8 U / mL. The relational expression between the SOD concentration and the adduct ratio for obtaining the SOD-like activity value of each material was SOD-like activity value (U / mL) = 1.119 × adduct ratio + 0.1053 (r = 0.996).

(Test Example 2) Measurement of SOD activity value of each material by ESR Antioxidant composition A obtained in Example 1, tea extract prepared and obtained in Comparative Example, oolong tea extract, grape seed extract and vitamin C Each sample was prepared by diluting with distilled water to 10 μg / mL. And it measured by the method similar to Test Example 1 and calculated | required the SOD-like activity value. The results are shown in Table 2.

  As shown in Table 2, the antioxidant composition A has a much higher SOD-like activity value than vitamin C, which is a general antioxidant, and a value much higher than the IC50 value. It was confirmed that there was an oxidizing effect.

(Example 2) Preparation 2 of an antioxidant composition
Amla dried fruit was pulverized to 40 mesh or less, 2 L of distilled water was added to 80 g of the powder, and extracted at 55 ° C. for 3 hours. Then, it centrifuged (3000 rpm, 10 minutes), the supernatant was filtered, and the extract and the residue were isolate | separated. Distilled water (2 L) was added to the residue, and extraction was repeated once more under the same conditions. The extracts were combined and concentrated under reduced pressure to 200 mL. Ethyl alcohol was added to this concentrated solution to prepare 1 L (final ethyl alcohol concentration 80%), and then allowed to stand at 4 ° C. for 24 hours, and then insoluble components were precipitated. The precipitate is separated and removed by centrifugation (3000 rpm, 10 minutes), the supernatant is concentrated under reduced pressure, redissolved in 1 L of water, filtered to remove insoluble components, the filtrate is freeze-dried, and the antioxidant composition of the present invention 12.5 g of product B was obtained.
Similarly, the final concentration of ethyl alcohol was 20%, 40%, and 60% to obtain 13.6 g, D20.8 g, and E21.2 g of the antioxidant composition of the present invention.

(Example 3) Preparation of antioxidant composition 3
Amla dried fruit was pulverized to 40 mesh or less, 2 L of distilled water was added to 80 g of the powder, and extracted at 55 ° C. for 3 hours. Then, it centrifuged (3000 rpm, 10 minutes), the supernatant was filtered, and the extract and the residue were isolate | separated. Distilled water (2 L) was added to the residue, and the mixture was repeatedly extracted under the same conditions. The extracts were combined and then lyophilized to obtain about 37.0 g of a dried product. 1 L of ethyl alcohol was added to 35 g of the dried product, and the mixture was allowed to stand at 4 ° C. for 24 hours to precipitate insoluble components. The precipitate is separated and removed by centrifugation (3000 rpm, 10 minutes), the supernatant is concentrated under reduced pressure, redissolved in 1 L of water, filtered to remove insoluble components, the filtrate is freeze-dried, and the antioxidant composition of the present invention 3.5 g of product F was obtained.

Example 4 Preparation 3 of Antioxidant Composition
Amlar dried fruit was pulverized to 40 mesh or less, 2 L of distilled water was added to 80 g of the powder, and extracted at 55 ° C. for 3 hours. Then, it centrifuged (3000 rpm, 10 minutes), the supernatant was filtered, and the extract and the residue were isolate | separated. Distilled water (2 L) was added to the residue, and extraction was repeated once more under the same conditions. The extracts were combined and concentrated under reduced pressure to 200 mL. Ethyl acetate was added to this concentrated solution to prepare 500 mL (final ethyl acetate concentration 60%), stirred well, allowed to stand at 4 ° C. for 24 hours, then the ethyl acetate layer was separated, concentrated under reduced pressure, The filtrate was lyophilized to obtain 12.5 g of the antioxidant composition G of the present invention.

(Test Example 3) Active oxygen inhibition measurement by ESR Antioxidant composition A obtained in Example 1, antioxidant compositions B, C, D and E obtained in Example 2, obtained in Example 3 Antioxidant composition F and antioxidant composition G obtained in Example 4 were each diluted with distilled water to 1 μg / mL to prepare a sample. And it measured by the method similar to Test Example 1 and calculated ^| required O2 ^- inhibition rate. Moreover, it melt | dissolved with ethyl alcohol and adjusted 30 mM DPPH (1,1dephenyl-2-picrylhydrazyl). Moreover, each antioxidant composition was newly diluted with ethyl alcohol so that it might become 1 microgram / mL, and the sample was adjusted. 100 μL of DPPH solution and 100 μL of sample were added to a test tube for ESR, mixed for 10 minutes, and then ESR signal was quantified under the same conditions as in Test Example 1 to determine DPPH free radical inhibition rate. These results are shown in Table 3.

As shown in Table 3, no significant difference was observed in the O ² -inhibitory effect even when the treatment was carried out at different ethyl alcohol concentrations. However, DPPH free radical inhibition was treated at an alcohol concentration of 60% or more. A high effect was observed in the antioxidant composition.

(Test Example 4) Preparation of STZ-induced diabetic nephropathy rat A test diet containing 24.0% casein, 3.5% lipid, and 60.5% carbohydrate was added to a Wistar male rat (Japan SLC, 8 weeks old). After pre-feeding for 1 week, STZ (50 mg / kg) was dissolved in citrate buffer and injected intraperitoneally. Thereafter, the feed was continuously fed until the test was completed. Rats injected intraperitoneally with STZ were collected from the tail vein one week later, and blood glucose levels and body weights were measured to confirm the production of diabetic nephropathy-induced rats.

(Test Example 5) Confirmation of in vivo antioxidant effect STZ-induced diabetic nephropathy rats were divided into 3 groups so that the average body weight was approximately 10 animals / group.
1) Normal group: normal rat group fed without injection of STZ for control 2) Control group: 0.5% physiological saline oral administration 3) Antioxidant composition A administration group: Antioxidant composition A) 20 mg / kg body weight / day at start of test administered orally 4) Antioxidant composition G administration group: antioxidant composition G 20 mg / kg of test body weight kg / day administered orally STZ-induced diabetic nephropathy rats group The composition was suspended in 0.5% physiological saline, orally administered daily for 20 days, and reared on the test feed of Test Example 4. The normal group was bred only with the test feed. Blood was collected before the test was completed, and the serum glucosylated protein value, serum creatinine value, and TBA reactive substance concentration were measured from the obtained blood based on a conventional method. The results are shown in Table 4.

＊ｐ＜０．０５対照に対して

* P <0.05 vs control

In vivo antioxidant effect As shown in Table 4, in the rats with STZ-induced diabetic nephropathy, the antioxidant composition-administered group had a lower TBA reactive substance concentration than the control group, and the peroxidation caused by the onset of diabetic nephropathy It was confirmed that the increase in lipid was suppressed by ingesting the antioxidant composition and the effect of improving diabetic nephropathy was also observed, and the antioxidant composition exhibited an in vivo antioxidant effect.

(Test Example 6) Lipid peroxide inhibition test of food The antioxidant composition E obtained in Example 2 and the antioxidant composition G obtained in Example 4 and α-tocopherol were added to 3 g of minced crab meat. Add 50 μL of ethyl alcohol prepared to 0.1 g / 10 mL, or 50 μL of ethyl alcohol (control) and mix well. These TBA reactant concentrations were measured by a conventional method after mixing and after storage at 4 ° C. for 4 days. Table 5 shows the ratio when the antioxidant composition was blended with the control TBA reactant concentration being 100%.

  As shown in Table 5, by adding an antioxidant composition to foods, it has a higher antioxidant effect that suppresses the reduction and production of lipid peroxides that cause food deterioration than commonly used α-tocopherol. Was recognized.

(Example 5) Preparation of antioxidant composition-containing food (tablet confectionery) Antioxidant composition A 5 g, lactose 30 g obtained in Example 1, DHA-containing powdered oil (Suncoat DY-5; manufactured by Taiyo Kagaku Co., Ltd.) 12 g, 4 g of sucrose fatty acid ester and 4 g of yoghurt flavor are mixed, and the mixture is pressure-molded using a rotary tableting machine, and 1 tablet is 300 mg of the antioxidant composition-containing food or drink of the present invention (tablet confectionery) Got.

Example 6 Preparation of Antioxidant Composition-Containing Beverage Antioxidant Composition B 5 g obtained in Example 2, 1/5 concentrated grapefruit clear fruit juice 2.1 g, erythritol 30 g, citric acid crystals 2.5 g, citric acid Trisodium 0.5g, L-ascorbic acid 0.5g, calcium lactate 1.93g, CCP 0.15g, grapefruit flavor 1.0 are mixed and dissolved in water to make a total volume of 1000ml, and it is filled into a 100ml bottle. After sealing with a cap, it was sterilized by heating at 90 ° C. for 30 minutes to obtain an antioxidant composition-containing food or drink according to the present invention.

Example 7 Preparation of Antioxidant Composition-Containing Beverage (Vegetable Juice Mixed Beverage) Antioxidant composition C1 g obtained in Example 2 and guar gum degradation product (Sunfiber R; manufactured by Taiyo Kagaku Co., Ltd.) 3 g are commercially available. The resulting mixture was dissolved in 100 ml of a mixed vegetable fruit juice to obtain an antioxidant composition-containing food or drink (vegetable juice mixed drink) of the present invention.

Example 8 Preparation of Antioxidant Composition-Containing Cookies After putting 4 g of the antioxidant composition D obtained in Example 2 and 200 g of commercially available cake mix powder into a container, add 35 g of butter and mix with a wooden coconut. It was. 25g of beaten egg was added to it and kneaded well until it became a smooth dough. The dough is taken out on a table on which the flour has been shaken, and the flour is further shaken and stretched to a thickness of 5 mm with a rolling pin, extracted with a round shape, and baked in an oven at 170 ° C. for 10 minutes, and about 5 g of this invention. An antioxidant composition-containing cookie was obtained.

(Example 9) Preparation of antioxidant composition-containing yogurt In Example 2, antioxidant composition E10 g, commercially available skim milk (manufactured by Meiji Dairies Co., Ltd., protein content 34%) 0.95 kg, and commercially available unsalted butter (Snow Brand) (Produced by Dairy Co., Ltd.) 0.35 kg was dissolved in 8 L of warm water, homogenized, and the total amount was adjusted to 10 L. Next, it is sterilized by heating at 90 ° C. for 15 minutes, cooled, inoculated with 0.03 kg of commercially available lactic acid bacteria starter (manufactured by Hansen) (0.02 kg of Streptococcus thermophilus and 0.01 kg of Lactobacillus bulgaricus), and mixed uniformly. Into a 100 ml container, filled, sealed, fermented at 37 ° C. for 20 hours, and then cooled to obtain the antioxidant composition-containing yogurt of the present invention.

(Example 10) Preparation of antioxidant composition-containing oral liquid food 100 g sodium caseinate (manufactured by DMV), 85 g egg white enzyme degradation product (manufactured by Taiyo Chemical Co., Ltd.), 200 g dextrin (manufactured by Matsutani Chemical Co., Ltd.) An aqueous phase was prepared in the tank. Separately, 90 g of MCT (manufactured by Kao Corporation), 35 g of palm oil (manufactured by Fuji Oil Co., Ltd.), 35 g of safflower oil (manufactured by Taiyo Oil & Fats Co., Ltd.), 1.4 g of lecithin (manufactured by Taiyo Chemical Co., Ltd.), antifoaming agent ( 2 g of Taiyo Kagaku Co., Ltd. was mixed and dissolved to prepare an oil phase. The oil phase was added to the aqueous phase in the tank, stirred and mixed, then heated to 70 ° C., and further homogenized with a homogenizer at a pressure of 14.7 MPa. Next, after sterilizing at 90 ° C. for 10 minutes, the solution was concentrated and spray-dried to prepare about 510 g of an intermediate product powder. 500 g of this intermediate product powder, 10 g of the antioxidant composition C obtained in Example 2, 390 g of dextrin (manufactured by Matsutani Chemical Co., Ltd.), 45 g of guar gum degradation product (Sunfiber R; manufactured by Taiyo Chemical Co., Ltd.), a small amount of vitamins and minerals and powder A fragrance was added and mixed uniformly to obtain about 950 g of an oral liquid food containing an antioxidant composition.

(Example 11) Preparation of antioxidant composition-containing tablet Antioxidant composition A (20 g) obtained in Example 1, 10 g of crystalline cellulose, 27.5 g of corn starch, 65 g of lactose, and 6.5 g of hydroxypropyl cellulose were mixed and granulated. Turned into. To this granulated product, 2.0 g of magnesium stearate is added, mixed uniformly, and this mixture is pressure-molded using a rotary tableting machine to obtain a tablet containing the antioxidant composition of the present invention, each containing 130 mg. It was.

Example 12 Preparation of Antioxidant Composition-Containing Capsule Antioxidant composition composition G10 g obtained in Example 4, 99 g of lactose, 9.4 g of corn starch, and 1.6 g of polyvinyl pyrrolidone are uniformly mixed to form a gelatin capsule The mixture was filled with 500 mg of this mixture to obtain an antioxidant composition-containing capsule.

Examples of the present invention and the target product are as follows.
(1) An antioxidant composition comprising Amler or an extract thereof.
(2) The antioxidant composition according to the above (1), wherein the extract of Amler is extracted from any of Amler fruit or fruit juice with water, base, acid, or hydrophilic solvent.
(3) The antioxidant composition as described in (1) or (2) above, wherein the extract of Amler is extracted from water or fruit juice with water.
(4) The description in (1) or (2), wherein the hydrophilic solvent is at least one lower alcohol selected from the group consisting of methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, and butyl alcohol. Antioxidant composition.
(5) The antioxidant composition according to any one of (1) to (4), wherein the antioxidant composition is fractionated from an extract of Amler fruit or fruit juice or fruit juice with an organic solvent.
(6) The organic solvent is at least selected from the group consisting of methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, butyl alcohol, ethyl acetate, butyl acetate, diethyl ether, methyl ether, methyl isobutyl ketone, hexane, or chloroform. The antioxidant composition according to (5) above, wherein the antioxidant composition is one of the following.
(7) The antioxidant composition according to any one of (1) to (6) above, wherein the extract is extracted with ethyl alcohol from an Amler fruit or fruit juice extract or fruit juice and is characterized by an ethyl alcohol-soluble fraction.
(8) The antioxidant composition according to (7) above, wherein the concentration of ethyl alcohol when fractionated with ethyl alcohol is 10 to 100% as a final concentration, and is an ethyl alcohol-solubilizing component.
(9) The concentration according to (7) or (8), wherein the concentration of ethyl alcohol when fractionating with ethyl alcohol is 60 to 100% as a final concentration and is an ethyl alcohol-solubilizing component. Oxidizing composition.
(10) The antioxidant composition according to any one of (1) to (5) above, wherein the extract is extracted with ethyl acetate from an Amler fruit or fruit juice extract or fruit juice and is characterized by an ethyl acetate-soluble fraction.
(11) The antioxidant composition according to the above (10), wherein the ethyl acetate concentration when fractionating with ethyl acetate is 20 to 80% as a final concentration, and is an ethyl acetate solubilizing component.
(12) The concentration according to (10) or (11) above, wherein the ethyl acetate concentration at the time of fractionation with ethyl acetate is 40 to 80% as a final concentration and is an ethyl acetate solubilizing component. Oxidizing composition.
(13) Food / beverage products containing the antioxidant composition in any one of said (1)-(12).
(14) A quasi-drug containing the antioxidant composition according to any one of (1) to (12).
(15) A pharmaceutical comprising the antioxidant composition according to any one of (1) to (12).
(16) A feed comprising the antioxidant composition according to any one of (1) to (12).

  The antioxidant composition containing the Amler extract obtained in the present invention has high active oxygen scavenging ability, SOD-like activity value and DPPH free radical scavenging ability in the test by the ESR method, and was induced by administering STZ. Antioxidant composition administration test to diabetic nephropathy rats and addition test to fish meat showed reduction and suppression of lipid peroxide TBARS, and high antioxidant effect even in vivo and food addition It can be confirmed that it can be used for various foods, quasi-drugs, pharmaceuticals, etc., aging, stroke, autoimmune disease, cancer, cataract, fatty liver, myocardial infarction, ischemic heart disease, Alzheimer's syndrome, diabetes, diabetes Various diseases such as nephropathy, arteriosclerosis, allergy, hay fever and abnormal pigmentation such as stains and freckles can be suppressed and food deterioration can be suppressed.

Claims (8)

An antioxidant composition comprising Amler or an extract thereof. The antioxidant composition according to claim 1, wherein the extract of Amler is extracted from Amler fruit or fruit juice with any of water, base, acid, hydrophilic solvent, and acetone. The antioxidant composition according to claim 1 or 2, wherein the hydrophilic solvent is at least one lower alcohol selected from the group consisting of methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, and butyl alcohol. The antioxidant composition according to any one of claims 1 to 3, which is fractionated with an organic solvent from an extract of Amler fruit or fruit juice, or fruit juice. The organic solvent is at least one selected from the group consisting of methyl alcohol, ethyl alcohol, n-propyl alcohol, isopropyl alcohol, butyl alcohol, ethyl acetate, butyl acetate, diethyl ether, methyl ether, methyl isobutyl ketone, hexane, or chloroform. The antioxidant composition according to claim 4, wherein the antioxidant composition is present. Food / beverage products containing the antioxidant composition in any one of Claims 1-5. A quasi-drug containing the antioxidant composition according to any one of claims 1 to 5. A pharmaceutical comprising the antioxidant composition according to any one of claims 1 to 5.