JP5282932B2 - Production method of polyphenol extract, osteoporosis preventive agent, carbohydrate digestive enzyme inhibitor, functional composition using these, and food composition, food composition for specified health use, quasi-drug containing this functional composition Product composition, pharmaceutical composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for producing a polyphenol extract containing no citric acid, having a well-defined polyphenol composition, and having a high polyphenol content; to provide a functional composition, a prophylactic agent for osteoporosis, a saccharide-digesting enzyme inhibitor for treating and preventing obesity and diabetes each using the polyphenol extract; and to provide a food composition, a food composition for specified health use, a quasi-drug composition, and a pharmaceutical composition. <P>SOLUTION: According to the polyphenol extract production method, a polyphenol extract containing no citric acid and high concentration of a polyphenol can be obtained by bringing ume plum vinegar into contact with a styrene/divinylbenzene-based synthetic adsorbent resin, subjecting the styrene/divinylbenzene-based synthetic adsorbent resin to elution treatment with an aqueous solvent, and concentrating the eluate. Further, the new functional composition of the polyphenol extract obtained from ume plum vinegar can be utilized in a food composition for diabetes prevention, a food composition for specified health use, a quasi-drug composition, and a pharmaceutical composition. <P>COPYRIGHT: (C)2009,JPO&amp;INPIT

Description

  The present invention relates to a method for producing a polyphenol extract obtained from ume vinegar, and a functional composition of a polyphenol extract obtained from ume vinegar. The present invention also provides an agent for preventing osteoporosis using a polyphenol extract obtained from ume vinegar, an α-amylase inhibitory action and an α-glucosidase inhibitory action effective for the treatment and prevention of obesity and diabetes. The present invention relates to a digestive enzyme inhibitor, a functional composition containing the same, and a food composition, a food composition for specified health use, a quasi-drug composition, and a pharmaceutical composition using the functional composition.

  In Japan, ume has been processed into plums, plum wine and plum extract since ancient times. Outside Japan, plums are eaten mainly on the Korean Peninsula, China and Taiwan. In China, the immature plum fruit is picked to obtain the Chinese herbal medicine “Plum”. “Saimei” is used for digestive diseases. In this way, plum is recognized as a healthy food.

  However, there has been little detailed analysis of the functional components of plums and human intervention studies. As a report regarding the functional component of plum, some reports have been made on, for example, polyphenols, pectic substances, or the glycosides of hydroxycinnamic acid in which plum flowers are present, such as pullose I, II, III.

  On the other hand, polyphenol has attracted attention as a functional material involved in the health function of fruits. Polyphenol is a general term for plant components having a plurality of phenolic hydroxyl groups (hydroxy groups) in the same molecule. Polyphenols are said to have antioxidative effects and are effective in preventing arteriosclerosis, senile dementia, cerebral infarction, rheumatic diseases, myocardial infarction, gout, diabetes, etc. .

  It is thought that polyphenols contained in plums are also involved in the health functions handed down in plums.

  For example, in umeshu, rioniresinol which is a lignan derivative which is a kind of polyphenol exists. It is known that this rioniresinol has an antioxidant action (for example, refer nonpatent literature 1).

  Also, syringalesinol, which is a kind of lignan derivative, has been isolated from plum. Syringalesinol has been reported to have an antibacterial action against Helicobacter pylori living in the gastric mucosa (see, for example, Non-Patent Document 2).

  Furthermore, it has been shown that an extract of plum fruit exhibits a blood pressure lowering effect in experimental animals (see, for example, Non-Patent Document 3). This document describes that chlorogenic acid, which is a kind of polyphenol contained in an extract of plum fruit, has an angiotensin converting enzyme inhibitory activity, which is an enzyme that converts angiotensin I into angiotensin II that increases blood pressure. ing.

  Furthermore, it has been reported that the extract of plum fruit suppresses the production of verotoxin at a concentration of about half the minimum growth element concentration against enterohemorrhagic Escherichia coli O157 (for example, non-patented). Reference 4). Active substances that inhibit verotoxin production have not been isolated, but are inferred to be polyphenols.

  However, plum is a fruit that cannot be eaten without processing. For this reason, various by-products and waste are generated in the processing process. The main by-product and waste is “plum vinegar” that is produced during the plum drying process. The amount of “plum vinegar” is about 16,000 tons per year in Wakayama Prefecture, for example, which calculates about 60% of Japanese umeboshi production.

  Conventionally, these ume vinegars have been used as an auxiliary material for vegetable pickles, or electrodialyzed to recover salt, and desalted ume vinegar has been used as a food material such as a fruit juice substitute. However, its use is only a part of the whole plum vinegar and its added value is low. In addition, it is necessary to treat ume vinegar that is not reused, such as incineration and sewage treatment. For this reason, it is required to effectively use ume vinegar with high addition.

  On the other hand, it is known that polyphenols are also contained in ume vinegar (see, for example, Non-Patent Documents 5 and 6).

  Non-Patent Document 5 discloses a method for obtaining a “plum vinegar extract” as follows. The ume vinegar is first filtered through diatomaceous earth to remove impurities, and then passed through a column of a synthetic adsorption resin (DIAION HP-20, manufactured by Mitsubishi Chemical Corporation). The resin is then washed with water to wash away salt and citric acid. This resin is washed with 90% ethanol of 1/20 volume of the amount of ume vinegar used, and the eluted liquid is concentrated to about 25% water to obtain a “plum vinegar extract”. This document discloses that this “plum vinegar extract” contains 0.3% of rioniresinol. In addition, it is disclosed that when this "plum vinegar extract" is administered to stroke-prone spontaneously hypertensive rats, it has an inhibitory effect on blood pressure, serum lipid peroxide, and liver neutral lipid accumulation. Has been.

  However, this document does not mention the polyphenol content and its type in the “plum vinegar extract”.

  Non-Patent Document 6 discloses “concentrated desalted plum vinegar” obtained by desalting plum vinegar by electrodialysis and then concentrating. This document describes that when this “concentrated desalted plum vinegar” is administered to hamsters, it has an effect of reducing the concentration of neutral lipids in serum and liver.

  However, the polyphenol content of this “concentrated desalted plum vinegar” is only 0.065%. Further, from the production method, it is presumed that this “concentrated desalted plum vinegar” contains a large amount of citric acid. Therefore, it is not clear whether the effect of the above-mentioned “concentrated desalted plum vinegar” is that of polyphenol. This may be the effect of a combination of polyphenols and citric acid.

  In order to effectively use plum vinegar, it is important to make an extract with a clear polyphenol composition and a high polyphenol content. In addition, by finding a new effect of polyphenol, a new field of use of ume vinegar can be developed.

  In recent years, attempts have been made to use plant extracts containing polyphenols as functional materials as health foods and pharmaceutical raw materials (see, for example, Patent Documents 1 and 2). For example, Patent Document 1 discloses an enzyme inhibitor using a plant extract of the family Ceratomyaceae. Further, Patent Document 2 discloses an α-amylase inhibitor using a petal part or fruit part extract of Hermanus.

  However, these plants have no food experience or use extremely poor materials. For this reason, a long time and cost are expected for safety evaluation. Polyphenol is a general term for complex compounds having a plurality of hydroxyl groups on the benzene ring, and there are a great number of types. Plant-derived polyphenols are different in the types of polyphenols contained in the plants and have different effects.

On the other hand, ume vinegar has been used and has been eaten. Ume vinegar, a by-product produced in the process of making plum pickles, is a traditional food widely used for pickles and seasonings, and its safety is not particularly problematic. Therefore, if a new function can be found in the polyphenol extract obtained from ume vinegar, its usefulness is high.
Shirasaka Noriaki, 7 others, “Antioxidation of Plum Wine and Isolation and Identification of Antioxidants”, Journal of Japan Society for Food Science and Technology, 1999, Vol. 46, p. 792-798 Otsuka T, et al., 9 others, “Suppressive effects of fruit-juice pumus pummus pulsium in the Mongolian gerbil of Mongolian gerbils. , Ume) on Helicobacter pylori-induced global stomach relations in Mongolian gerbils.), “Asia Pacific Journal of Cancer Prevention (Asian Pacific Journal 5) p . 337-341 Ina H (Ina H) and three others, “Effects of benzyl glucoside and effects of ume-derived benzyl glycoside and chlorogenic acid on plasma corticosteroid (ACTH) and catecholamine levels in experimental menopause model rats chlorogenic acid from Prunus mume on adrenocorticotropic hormone (ACTH) and catecholamine levels in plasma of experimental menopausal model rats) ", 2004, biological and Pharmaceutical Bulletin (bIOLOGICAL & PHARMACEUTICAL BULLETIN), Institute of the Pharmaceutical Society of Japan 2004, Vol. 27, p. 136-137 Sakagami Y, “Inhibitory Effect of the Extraction of Protractor Protochemium Ecitoh. In Zuccc. On Zercc. : H7) ", Biocontrol Science, Japanese Society for Antifungal Prevention, 2001, Vol. 6, p. 53-56 Tsuneji Kodera and 4 others, “Effect of ume vinegar extract on blood pressure and lipid metabolism in spontaneously hypertensive rats with high cholesterol diets”, Journal of Japanese Society of Nutrition and Food, Japanese Society of Nutrition and Food, 2004 57, p. 249-255 Tomohiro Tanaka and three others, “Effects of concentrated desalted plum vinegar on lipid metabolism in hamsters”, “Trace Nutrients Research”, Micronutrients Study Group, 2005, Vol. 22, p. 131-134 Patent Publication 2004-168770 Patent Publication 2005-306801

  That is, the present invention has been made in view of the above problems, and an object thereof is to produce a polyphenol extract that does not contain citric acid, has a clear polyphenol composition, and has a high polyphenol content. .

  Another object of the present invention is to provide a functional composition using a polyphenol extract having a high polyphenol content. Moreover, it exists in finding the new function of the polyphenol obtained from the ume vinegar using the extract with much content of polyphenol.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have made ume vinegar contact with a styrene-divinylbenzene synthetic adsorption resin, and then eluted with an aqueous solvent into the styrene-divinylbenzene synthetic adsorption resin. By concentrating the treated material, it was found that a polyphenol extract not containing citric acid and containing a high concentration of polyphenol was obtained, and the present invention was completed.

  Moreover, the novel function was discovered in the polyphenol extract obtained from ume vinegar. That is, the present invention is as follows.

  The method for producing a polyphenol extract of the present invention comprises a step of bringing ume vinegar into contact with a styrene-divinylbenzene synthetic adsorption resin, a step of washing the styrene-divinylbenzene synthetic adsorption resin with water, and the styrene-divinylbenzene series. A step of elution treatment of the synthetic adsorption resin with an aqueous solvent and a step of concentrating the eluate obtained by the elution treatment are included.

  Moreover, the functional composition of this invention is good to contain the polyphenol extract obtained by the said manufacturing method, or one part.

  The polyphenol extract obtained from ume vinegar functions as an osteoporosis preventive agent, an α-glucosidase inhibitor, and an α-amylase inhibitor. Moreover, the functional composition of the present invention can be used as a food composition, a food composition for specified health use, a quasi-drug composition, and a pharmaceutical composition.

  In the method for producing a polyphenol extract of the present invention, ume vinegar polyphenol is adsorbed on an adsorption resin. Citric acid and sodium chloride can be easily removed by washing with water. Moreover, if the component which was not adsorb | sucked to resin is collect | recovered, it can be reused for food etc. as it is.

  An aqueous solvent is used as the elution solvent. As a result, the polyphenol recovery rate can be increased. Moreover, since the aqueous solvent to be used does not contain water, the solvent can be easily removed.

  The polyphenol extract of the present invention contains almost 100% polyphenol. As a result, when this polyphenol extract is used, it can be effectively used as a functional composition using polyphenol. Moreover, what contains a part of polyphenol extract which isolate | separated desired polyphenol from the multiple types of polyphenol which comprises the polyphenol extract of this invention can be effectively utilized as a functional composition using polyphenol.

  In this invention, it discovered that the polyphenol extract obtained from plum vinegar had an osteoporosis prevention effect, (alpha) -amylase, or (alpha) -glucosidase inhibitory effect. As a result, the polyphenol extract of the present invention has an effect of preventing diabetes due to an osteoporosis preventive agent and a hypoglycemic action. That is, the functional composition of the present invention can be used for a food composition for preventing osteoporosis or for preventing diabetes, a food composition for specific health use, a quasi-drug composition, and a pharmaceutical composition.

  The present invention is described in detail below.

[Production method of polyphenol extract]
(Raw materials used)
In the present invention, ume vinegar is the raw material for the polyphenol extract. In the present specification, “plum vinegar” is produced in a step of salting plum fruits, and is composed of salt and a solution leached from plum fruits. Generally, ume vinegar contains about 20% by mass of salt.

  The fruit of Minamitakaume, which is a typical plum variety, contains about 9000 ppm of polyphenols, depending on the time of collection. The main polyphenol is hydroxycinnamic acid glycoside. However, it is difficult to extract a sufficient amount of polyphenols from plum fruit.

  On the other hand, the ume vinegar 1t contains about 30 g of polyphenol. Moreover, the composition of the polyphenol is the same as that of the polyphenol contained in the fruit of a plum.

(Adsorption resin)
As the adsorption resin to be used, an adsorbent capable of selectively adsorbing and eluting polyphenols, for example, a styrene-divinylbenzene synthetic adsorption resin may be used. Specifically, Diaion (registered trademark) HP20, HP21 (manufactured by Mitsubishi Chemical Corporation), Sepabeads (registered trademark) SP825L, SP850 (Mitsubishi Chemical) are synthetic adsorption resins having a porous structure having no ion exchange group. Sepa Beads (registered trademark) SP700 (manufactured by Mitsubishi Chemical Corporation), Sepa Beads (registered trademark) SP70 (manufactured by Mitsubishi Chemical Corporation), and the like can be used. Particularly preferred resins are Diaion (registered trademark) HP20 and Sepabeads (registered trademark) SP70. When these resins are used, about 90% of polyphenols in ume vinegar can be adsorbed on the resin.

(Elution solvent)
The elution of polyphenol from the styrene-divinylbenzene synthetic adsorption resin is performed with an aqueous solvent. In this specification, an aqueous solvent is an organic solvent having an affinity for water, and examples thereof include alcohols having 1 to 5 carbon atoms and acetone. Ethanol and acetone are preferred. Depending on the styrene-divinylbenzene synthetic adsorption resin used, about 70% or more of the polyphenol contained in ume vinegar can be recovered using ethanol, and nearly 90% of the polyphenol contained in ume vinegar can be recovered using acetone. These aqueous solvents are preferably not used as an aqueous solution, but as an aqueous solvent alone or a mixture of aqueous solvents.

(Production method)
First, ume vinegar is brought into contact with a styrene-divinylbenzene synthetic adsorption resin, and polyphenols in ume vinegar are adsorbed onto the styrene-divinylbenzene synthetic adsorption resin. As the styrene-divinylbenzene synthetic adsorption resin, an amount having an allowance for adsorbing polyphenol contained in ume vinegar is used. What is necessary is just to investigate beforehand the adsorption capacity of the resin to be used. For example, a styrene-divinylbenzene synthetic adsorption resin 20 times as much as ume vinegar is used.

  Moreover, you may collect | recover the component which was not adsorb | sucked in the styrene- divinylbenzene type synthetic adsorption resin in this case. The ume vinegar flavor remains in the components not adsorbed on the styrene-divinylbenzene synthetic adsorption resin. Therefore, it can be reused as it is for food.

  Next, the styrene-divinylbenzene synthetic adsorption resin adsorbed with polyphenol is washed with water. Thereby, the citric acid and salt contained in ume vinegar are removed from the styrene-divinylbenzene synthetic adsorption resin.

  Next, the styrene-divinylbenzene synthetic adsorption resin after the washing step is subjected to elution treatment with an aqueous solvent. Thereby, the polyphenol adsorbed on the resin can be recovered with high efficiency.

  Next, the eluate is concentrated. In the present invention, the eluate is an aqueous solvent containing no water and having a low molecular weight. Therefore, the solvent can be removed at a relatively low temperature. As a result, the energy for removing the solvent can be suppressed. The polyphenol extract thus obtained is in a viscous extract state.

  The above polyphenol extract may be produced by any one of the following batch method and continuous method in consideration of the amount of treatment.

  In the batch method, ume vinegar and an adsorbent resin are mixed in a large container, and the polyphenol in the ume vinegar is adsorbed on the adsorbent resin. Thereafter, the adsorption resin is washed with water. After washing, an aqueous solvent is added to elute the polyphenol. This method is inferior in polyphenol recovery and increases the amount of work. However, when a relatively small amount of plum vinegar within 1 ton is processed, no special production equipment is required, which is a useful method.

  The continuous method is a column method in which an adsorption resin is packed in a column. Plum vinegar is passed through a column packed with adsorption resin. Next, water is passed through the column to wash the adsorbent resin. After washing, an aqueous solvent is passed through to elute the polyphenol. This method is more efficient and enables a large amount of processing by appropriately designing conditions such as adsorption temperature, flow rate, and column shape. Also, labor saving and automation can be achieved. Moreover, the collection | recovery of the ume vinegar after polyphenol elution is also easy.

[Polyphenol extract]
The polyphenol extract of the present invention obtained by the above method usually contains 11% by mass or more of polyphenol in the extract. Polyphenols usually exist in the form of glycosides. The content of the polyphenol is the content of polyphenol that does not contain sugar. Polyphenols can be quantified by the foreign-thiocult method. Moreover, the polyphenol extract of the present invention does not contain citric acid.

  The polyphenol extract of the present invention may contain a part of this polyphenol extract. The polyphenol extract of the present invention contains many types of polyphenols. Among these, a polyphenol used for a specific purpose may be subjected to a sorting process such as filtration, column treatment, and solvent washing.

  There is no particular limitation on the form of the polyphenol extract of the present invention or a part of this extract, and the functional composition containing these, and it may be a solution or a solid, and may be a mixture with other compounds. There may be. In the case of a mixture, a mixture obtained by adding a shaping agent such as spray drying, freeze drying, or dextrin may be used.

  The functional composition of the present invention can be used as it is, but if necessary, it is used in pharmaceuticals, quasi drugs, cosmetics, bath preparations, foods, beverages and the like shown below. Any of the various components and additives that are suitable for the application can be arbitrarily selected and used in combination so that each product can be easily used.

  Specific examples of the functional composition of the present invention include polyphenols conventionally effective, anti-carcinogenic activity, arteriosclerosis, hypertension, senile dementia, cerebral infarction, rheumatic disease, myocardial infarction, gout, It can be used as a preventive agent such as diabetes, an antioxidant, and an antibacterial agent. Moreover, the polyphenol extract of this invention or a part of this extract can also be used as a tool which verifies the effect of the new polyphenol which is not known now. In the present invention, the polyphenol extract of ume vinegar has an effect of increasing bone density, and has found a new function as an osteoporosis preventive agent. Moreover, the function as an alpha-glucosidase inhibitor or an alpha-amylase inhibitor was discovered in the polyphenol extract of the ume vinegar.

  The effect as an osteoporosis preventive agent is to actually measure the bone density and confirm that the bone density increases.

  The α-glucosidase inhibitor or α-amylase inhibitor of the present invention contains a polyphenol extract obtained from the ume vinegar. The α-glucosidase inhibitory activity or α-amylase inhibitory activity is measured as follows.

(Measurement of α-glucosidase inhibitory activity)
The amount of p-nitrophenol produced by the reaction of α-glucosidase derived from rat small intestine acetone extract and p-nitrophenyl-α-D-glucopyranoside was measured colorimetrically, and extracted with ume vinegar-derived polyphenol The α-glucosidase inhibitory activity of the product is measured.

(Measurement of α-amylase inhibitory activity)
The reaction of porcine pancreatic α-amylase and soluble starch, and the reducing end such as maltose produced by hydrolysis of starch was quantified with parahydroxybenzene hydrazide (PAHBAH), and the polyphenol extract derived from plum vinegar The α-amylase inhibitory activity is measured.

  The usage-amount of the functional composition of this invention can be suitably changed according to the objective to be used. For example, when used in a pharmaceutical composition, an amount having an activity effect equivalent to that of a known α-glucosidase inhibitor may be used. Moreover, when using it for the food composition for diabetes prevention, the food composition for specific health, etc., the amount of use is reduced more than that. Moreover, when utilizing as an osteoporosis preventive agent, the quantity which a bone density amount actually increases is used.

  When the functional composition of the present invention is used for a pharmaceutical product or quasi drug, it can be administered orally. The perilla extract can be formulated with a pharmaceutical carrier and used in the form of tablets, capsules, drinks, troches, solutions, granules, powders, and the like. In that case, conventional ingredients such as excipients, lubricants, binders, disintegrants, solvents, suspending agents, stabilizers, colorants, sweeteners and the like can be added as appropriate.

  Other antiseptics for sprays, suppositories, preservatives, polishes, soaps, hygiene products, ointments, skin adhesive films, scratches, cuts, burns, scratches, inflammation, eczema, pimples, acne, rough skin, etc. Or a medical aid for treatment or a gauze impregnated with it, a mask, a bandage, an eye care agent, an inhalant, an oral cleansing agent, a mouthwash, a toothpaste and the like.

  For example, when ingested orally, it can be ingested by adding to food as a food additive. When used as a food additive, the amount added is not particularly limited, and may be appropriately determined according to the type of food. For example, liquid foods such as soft drinks and carbonated drinks, and solid foods such as confectionery and other various foods can be added and used. In addition, an example of an administration method when administered to the human body as food is as follows. Administration can be performed by various methods, for example, oral administration using tablets, capsules, granules, syrups and the like. The oral preparation can be produced according to a normal production method. For example, excipients such as starch, lactose and mannitol, binders such as sodium carboxymethyl cellulose and hydroxypropyl cellulose, disintegrants such as crystalline cellulose and carboxymethyl cellulose calcium, talc, stearin It can be manufactured as tablets, capsules, granules, and the like by appropriately combining a lubricant such as magnesium acid and a fluidity improver such as light anhydrous silicic acid.

  In the case of ingesting parenterally, application to the skin, spraying and the like can be mentioned, which can be used by adding to cosmetics. The addition amount is not particularly limited, and may be appropriately determined according to the type of cosmetic.

  Examples of cosmetics include lotions, emulsions, creams, ointments, shaving mousses, oils, packs, shampoos, rinses, treatments, hair tonics, hair styling agents, hair restorers, Skin care and hair care preparations such as permanent liquids, hair dyes, body soaps, or bath preparations (any form such as liquid, powder, granule, solid, etc.). .

  Specific examples of the use of the functional composition of the present invention for food include fishery processed products such as kamaboko, chikuwa and hampen, processed meat products such as sausage, ham and wine, tofu, fried chicken, konjac and the like Processed agricultural products, Western confectionery, Japanese confectionery, bread, cake, jelly, pudding, snacks, cookie, gum, candy, ramune and other confectionery, raw noodles, Chinese noodles, buckwheat, noodles and other noodles, sauces, Seasonings such as soy sauce, dressing, mayonnaise, sauce, honey, powdered candy, syrup, etc., spices such as curry powder, mustard powder, pepper powder, jam, marmalade, chocolate spread, pickles, Soy vegetables, sprinkles, processed vegetables and fruits such as canned and bottled vegetables and fruits, dairy products such as cheese, butter and yogurt, fruit juice, vegetable juice, whey drink , Soft drinks, health , Drinks such as medicinal liquor, and other health-oriented foods and drinks such as tablets, beverages, granules, etc., livestock feed, pet food etc. .

  The functional composition of the present invention may use additives generally used for granulation, such as excipients, binders, disintegrants, lubricants, talc, and fluidity improvers. For example, starch cellulose, lactose, crystalline cellulose such as mannitol carboxymethyl cellulose, hydroxypropyl cellulose, light anhydrous silicic acid such as carboxymethyl cellulose calcium, magnesium stearate, cellulose acetate, Chitosan, calcium alginate, calcium alginate, defatted rice sugar, oil cake, soy flour, wheat flour, anhydrous silicic acid, bran, rice flour, calcium carbonate, milk calcium, magnesium oxide, sodium bicarbonate, crystalline cellulose, starch, By formulating beer yeast, sugar, reduced lactose, vegetable oil and the like in combination as appropriate, it can be produced in the form of tablets, capsules, granules and the like.

  In addition, various additives derived from plant or animal raw materials and other additives can be used in combination as required. These additives are processed in a conventional manner according to the product type and form to be added (for example, pulverization, milling, washing, hydrolysis, fermentation, purification, pressing, extraction, fractionation, filtration, drying) , Powdering, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc., may be arbitrarily selected and combined), and may be selected arbitrarily from various materials. In addition, what is necessary is just to select the solvent used for extraction in consideration of the usage of the product to be provided (food use, external use, bath use), processing to be performed later, and the like.

  The functional composition of the present invention includes other moisturizers, hormones, sequestering agents, pH adjusters, chelating agents, antiseptic / antibacterial agents, cooling agents, stabilizers, emulsifiers, animal / plant properties. Proteins and degradation products thereof, animal and plant polysaccharides and degradation products thereof, animal and plant glycoproteins and degradation products thereof, blood flow promoters, anti-inflammatory agents, antiallergic agents, cell activators, keratolytic agents, wounds A therapeutic agent, a foaming agent, a thickener, an oral preparation, a deodorant / deodorant, a bittering agent, a seasoning, an enzyme, and the like may be used in combination. By combined use with these, additive and synergistic effects can be expected.

  Hereinafter, the present invention will be described in detail with reference to experimental examples, but the present invention is not limited to the experimental examples.

Example 1
In a container, 20 times amount (mass ratio) plum vinegar (polyphenol 270 mg / Kg) was contacted with a synthetic adsorption resin (Diaion (registered trademark) HP-20, manufactured by Mitsubishi Chemical Corporation). After removing the plum vinegar, DIAION HP-20 was washed with 5 times the amount of resin (mass ratio) of water. The amount of polyphenols in the ume vinegar that passed through the resin was quantified by the foreign-thiocult method. From the removed ume vinegar, it was found that about 90% by mass of the polyphenol of ume vinegar was adsorbed.

  Next, the adsorption resin to which the polyphenol was adsorbed was treated with methanol as an eluent to recover the polyphenol. The amount of methanol used was 5 times the amount of resin (mass ratio). The obtained polyphenol was about 65% by mass of the polyphenol of the original plum vinegar. The acidity of the obtained polyphenol extract was titrated. The obtained polyphenol extract did not contain citric acid.

(Example 2)
The procedure was the same as Example 1 except that ethanol was used as the eluent (5 times the amount of resin (mass ratio)). The obtained polyphenol was about 70% by mass of the original ume vinegar polyphenol. Further, citric acid was not contained in the obtained polyphenol extract.

(Example 3)
The same procedure as in Example 1 was carried out except that acetone was used as the eluent (5 times the amount of resin (mass ratio)). The obtained polyphenol was about 80% by mass of the original ume vinegar polyphenol. Further, citric acid was not contained in the obtained polyphenol extract.

Example 4
A synthetic adsorption resin (Separbeads (registered trademark) SP-70, manufactured by Mitsubishi Chemical Corporation) was used in place of the synthetic adsorption resin (Diaion (registered trademark) HP-20, manufactured by Mitsubishi Chemical Corporation). In the same manner as in Example 1. The obtained polyphenol was 70% by mass of the original ume vinegar polyphenol. Further, citric acid was not contained in the obtained polyphenol extract.

(Example 5)
A synthetic adsorption resin (Separbeads (registered trademark) SP-70, manufactured by Mitsubishi Chemical Corporation) was used as in Example 1 except that ethanol was used as the eluent (5 times the amount of resin (mass ratio)). I made it. The obtained polyphenol was 75% by mass of the polyphenol of the original plum vinegar. Further, citric acid was not contained in the obtained polyphenol extract.

(Example 6)
A synthetic adsorption resin (Separbeads (registered trademark) SP-70, manufactured by Mitsubishi Chemical Corporation) was used, and acetone was used as an eluent (5 times the amount of resin (mass ratio)). I made it. The obtained polyphenol was about 90% by mass of the polyphenol of the original plum vinegar. Further, citric acid was not contained in the obtained polyphenol extract.

(Comparative Example 1)
Using a synthetic adsorption resin (Separbeads (registered trademark) SP-70, manufactured by Mitsubishi Chemical Corporation), as an eluent, 90 volume% ethanol, 50 volume% ethanol, 90 volume% acetone, 50 volume% acetone were used. Except for this, the procedure was the same as in Example 1. The obtained polyphenol is the original ume vinegar polyphenol, about 90% by mass in the case of 90% by volume ethanol, about 60% by mass in the case of 50% by volume ethanol, about 80% by mass in the case of 90% by volume acetone, 50% In the case of volume% acetone, it was about 70 mass%.

  Thus, it can be seen that when a styrene-divinylbenzene synthetic adsorption resin is used and an aqueous solvent not containing water is used, the polyphenol recovery rate is good. In particular, ethanol (Diaion HP-20: about 70% by mass, Sepabead SP70: about 75% by mass), acetone (Diaion HP-20: about 80% by mass, Sepabead SP70: about 90% by mass), and extremely polyphenols. It can be seen that the recovery is excellent.

(Experimental example 1)
(Animal used)
An osteoporosis model rat was prepared as follows. SPF Wister female rats (13 weeks of age at the time of introduction) were allowed to acclimatize for several days, and then ovariectomy (OVX) and sham operation (sham) were performed. After that, those with a recovery period were used.

(feed)
As a control feed, the total amount of 1 kg of sucrose 585 g, casein 200 g, cellulose 100 g, corn oil 70 g, mineral mixture (AIN-93M, nutrition using mice and rats published in 1993 by the National Institute of Nutrition (AIN) 35 g standard purified feed for research) and 10 g vitamin mixture (AIN-93, standard purified vitamin mixture for nutritional studies using mice and rats published in 1993 by the National Institute of Nutrition (AIN)) Feed was used.

  Plum vinegar polyphenol-added feed (plum vinegar p.p-added feed) was prepared by adding 25 g of “plum vinegar polyphenol” containing 15% of the polyphenol obtained from the above examples, with the amount of sucrose in the control feed being 560 g. .

(Breeding method)
The rats after surgery were divided into the following 4 categories (5 per group) and reared for 28 days. Body weight and food intake were measured during the feed period.
1st section: used rat: sham, breeding feed: control feed 2nd section: used rat: sham, rearing feed: plum vinegar p. p-added feed 3rd section: used rat: OVX, rearing feed: control feed 4th section: used rat: OVX, rearing feed: plum vinegar p. p-added feed

(Evaluation)
After the feeding period, the animals were fasted for 24 hours. Thereafter, the femur was collected, and the bone density was measured by X-ray CT for laboratory animals manufactured by Aloka. The results are shown in FIG.

FIG. 1 shows each of the four groups (sham + control feed group (1 group), sham + plum vinegar pp-added feed group (2 wards), OVX + control feed group (3 wards), OVX + plum vinegar pp-added feed group (4 ³ ) represents the total bone density (mg / cm ³ ) of the rats in each group. In FIG. 1, sham + plum vinegar p. p-added feed group, OVX + control feed group, OVX + plum vinegar p. The p-added feed group was significant with a significant difference p <0.01. From FIG. 1, even in rats subjected to sham surgery (sham), the ume vinegar p.p. was compared to the bone density (741.0) of rats fed the control diet. It can be seen that the bone density amount (753.0) of the rats fed with the p-added diet increased. In rats subjected to ovariectomy surgery (OVX), the ume vinegar p.p. was compared to the bone density (691.0) of rats fed the control diet. It can be seen that the bone density amount (723.0) of the rats fed with p-added diet increased significantly. That is, it has been found that ume vinegar polyphenol has an effect of suppressing a decrease in bone mass.

  From the body weight and the amount of food intake, it is presumed that rats subjected to ovarian removal surgery (OVX) ingested an average of 33 mg / kg of ume vinegar polyphenol per day. The bone density recovery effect of this ume vinegar polyphenol was comparable to the results of the recovery test performed with the same amount of isoflavone.

  From the above, it can be seen that ume vinegar polyphenols are effective as a preventive agent for osteoporosis.

  The polyphenol extract derived from ume vinegar containing 15% of the polyphenol obtained from the above examples was used in the following experiment.

(Experimental example 2)
250 mg of rat small intestine acetone extract (manufactured by Sigma) was suspended in 5 ml of 100 mM phosphate buffer (pH 6.8) and centrifuged at 3,000 rpm. The supernatant was separated and filtered through a glass filter to obtain an α-glucosidase enzyme solution. 40 μl of α-glucosidase enzyme solution and 1,000 μl of sample solution in which polyphenol extract derived from ume vinegar was dissolved in 100 mM phosphate buffer (pH 6.8) at various concentrations were mixed, and the mixture was incubated at 37 ° C. for 5 minutes. Incubated. Next, 950 μl of 0.7 mM p-nitrophenyl-α-D-glucopyranoside (manufactured by Sigma) and 100 mM potassium phosphate buffer ((pH 6.8), dissolved in 100 mM NaCl) are mixed, and 37 ° C. for 15 minutes. Incubated. Thereafter, 1 ml of a reaction stop solution (0.5 M Tris solution) was added, and the absorbance was measured at 400 nm. Separately, distilled water was used instead of the sample solution as a control, and before adding a p-nitrophenyl-α-D-glucopyranoside solution as a blank, a reaction stop solution was added and the same operation was carried out to measure the absorbance. The results are shown in FIG.

  In FIG. 2, the horizontal axis represents the concentration (μg / ml) of the polyphenol extract derived from ume vinegar added to the reaction solution, and the vertical axis represents the absorbance when no polyphenol extract derived from ume vinegar is added. The percentage of relative activity when expressed as% is expressed in%. It has been found that when the polyphenol extract derived from ume vinegar exceeds 500 μg / ml, α-glucosidase is inhibited in a concentration-dependent manner. It has been found that a polyphenol extract derived from ume vinegar inhibits α-glucosidase in a concentration-dependent manner.

(Experimental example 3)
Porcine pancreatic α-amylase (manufactured by Sigma) was dissolved in artificial saliva to obtain an enzyme solution. Soluble starch (manufactured by Nacalaisk) was suspended with artificial saliva to 1%, and dissolved by treatment at 100 ° C. for 15 minutes. Artificial saliva was prepared by the method described in the literature (Gal, J. Y., et. Al., About a synthetic saliva for in vitro studies. Talanta (2001), 53, 1103-1115). 100 μl of α-amylase solution and 900 μl of artificial saliva containing polyphenol extracts derived from ume vinegar with various concentrations were mixed and incubated at 37 ° C. for 5 minutes. Next, 500 μl of a soluble starch solution was added, and further incubated at 37 ° C. for 25 minutes. 1 ml of a PAHBAH solution prepared by mixing PAHBAH (manufactured by Sigma) in a 0.5 M HCl solution at a concentration of 5% and a 0.5 M NaOH solution in a ratio of 1: 4, and 50 μl of the above reaction solution Were mixed and heated at 100 ° C. for 10 minutes. The absorbance of this reaction solution was measured at 410 nm. A blank containing no enzyme was used. The results are shown in FIG.

  In FIG. 3, the horizontal axis represents the concentration (μg / ml) of the polyphenol extract derived from ume vinegar added to the reaction solution, and the vertical axis represents the absorbance when the polyphenol extract derived from ume vinegar is not added. The percentage of relative activity when expressed as% is expressed in%. It was found that the polyphenol extract derived from ume vinegar inhibits α-amylase in a concentration-dependent manner.

  From the above, it can be seen that the polyphenol extract derived from ume vinegar inhibits carbohydrate digestive enzymes, and thus suppresses an increase in blood glucose level after meals and is effective as a prophylactic agent for diabetes.

FIG. 1 is a graph showing the total bone density in rats in each experimental group. FIG. 2 is a diagram showing that a polyphenol extract derived from ume vinegar inhibits α-glucosidase. FIG. 3 is a diagram showing that a polyphenol extract derived from ume vinegar inhibits α-amylase.

Claims (7)

A step of bringing the ume vinegar into contact with a styrene-divinylbenzene synthetic adsorption resin packed in a column ;
Washing the styrene-divinylbenzene synthetic adsorption resin with water;
Elution treatment of the styrene-divinylbenzene synthetic adsorption resin with an aqueous solvent not containing water ;
And a step of concentrating the eluate obtained by the elution treatment, to produce a polyphenol extract containing no citric acid .   The functional composition containing the polyphenol extract obtained by the manufacturing method of Claim 1.   The functional composition containing a part of polyphenol extract obtained by the manufacturing method of Claim 1. Containing polyphenols Lumpur extract obtained from vinegar, osteoporosis prophylactic agent. A step of bringing the ume vinegar into contact with a styrene-divinylbenzene synthetic adsorption resin packed in a column;
Washing the styrene-divinylbenzene synthetic adsorption resin with water;
Elution treatment of the styrene-divinylbenzene synthetic adsorption resin with an aqueous solvent not containing water;
A step of concentrating the eluate obtained by the elution treatment;
Including, including polyphenols Lumpur extract obtained by the method for producing a polyphenol extract containing no citric acid, alpha-glucosidase peptidase inhibitors and alpha-amylase inhibitor. A functional composition having the function according to claim 4 or 5 . A food composition, a food composition for specified health use, a quasi-drug composition, and a pharmaceutical composition comprising the functional composition according to claim 6 .

Images