KR20060069885A - Process for producing product containing proanthocyanidin in high proportion - Google Patents

Abstract

Easily obtaining a proanthocyanidin-containing product wherein OPC of high physiological activity is contained in high proportion by treating a plant extract or juice with at least two types of adsorbents differing in at least one of material, pore radius, specific surface area and molecular weight distribution range.

Description

Process for producing product containing proanthocyanidin in high proportion

The present invention relates to a method for efficiently preparing a proanthocyanidin-containing content containing a large amount of physiologically active OPC.

Proanthocyanidins are condensation or polymerization of tannins present in various plants (hereinafter referred to as condensation polymerization), and compounds condensation-polymerized using flavan-3-ol or flavan-3,4-diol as structural units It's a military. These are given the name proanthocyanidin in that they produce anthocyanidins, such as cyanidin, delphinidin, and pelagonidine, by acid treatment.

Proanthocyanidins are a type of polyphenol, a potent antioxidant produced by plants, and concentrated in the leaves, bark, fruit bark or seeds of plants. Proanthocyanidins are specifically contained in grape seeds, pine bark, peanut bark, ginkgo biloba, acacia, bilberry. It is also known that proanthocyanidins are also contained in West African colanuts, Peruvian ratania roots, and Japanese green tea. Proanthocyanidins are substances that cannot be produced in the human body.

Proanthocyanidins are generally obtained by extraction from plants. As a solvent used for extraction, it is water; Organic solvents such as methanol, ethanol, acetone, hexane and ethyl acetate; Or mixtures thereof (Japanese Patent Application Laid-Open No. 11-80148). However, simply by solvent extraction, the recovery amount of proanthocyanidins is low and the purity is low. Therefore, in order to use as a raw material for health foods, cosmetics and pharmaceuticals, a process such as new concentration, purification, etc. is required to increase the purity, and it takes cost and time.

A method for recovering polyphenols containing proanthocyanidins has been reported. For example, Japanese Patent Application Laid-Open Nos. 5-279264 and 6-56689 disclose that polyphenols are adsorbed to chitin to use chitin with polyphenols adsorbed as a polyphenol product. Patent Publication No. 2002-97187 discloses free poly by adding ascorbic acid and an alkali metal or a salt thereof to a plant extract, precipitating a metal salt of polyphenols with a pH of 6 to 11, and desalting such a precipitate with an ion exchange resin or the like. A method for recovering phenol is described.

By the way, in recent years, especially among proanthocyanidins, the condensation polymer (2-4 tetramer) of the polymerization degree 2-4 is reported to be excellent in antioxidant activity. In the present specification, a condensate having a degree of polymerization of 2 to 4 is referred to as OPC (oligomeric proanthocyanidin). OPC also has the effect of reducing plaque (tartar) by inhibiting bacterial growth in the oral cavity in addition to antioxidant activity; Effect of restoring elasticity of blood vessels; Preventing lipoproteins from being damaged by free radicals in the blood, thereby causing the damaged lipoproteins to aggregate on the inner wall of blood vessels and preventing cholesterol from adhering to them; Effect of regenerating vitamin E degraded by free radicals; It is known to have an effect as an enhancer of vitamin E and the like.

However, most of the proanthocyanidins recovered by the above-described method have a high degree of polymerization, and the content of OPCs of 2- to 4-mers having high physiological activity is very low.

Japanese Patent Application Laid-Open Nos. 4-190774, 10-218769, 2001-131027, and Eberhard Scholz et al., Proanthocyanidins from Krameria triandra Root, Planta Medica, 55 (1989) ), Pages 379-384, describe how to extract OPC from plants and / or how to synthesize OPC. However, in the extraction method, the extract obtained by contacting the extract of the plant with the adsorbent, eluting the adsorbed components is recovered, and then using the fraction, the OPC content having a high OPC content is not repeated. It cannot be obtained and is not efficient. Also in the synthesis method, there are many processes, such as cost and time. There is also a problem of waste liquid treatment.

Therefore, there is a need for a method for purifying proanthocyanidins containing a large amount of OPC.

Disclosure of the Invention

The present inventors earnestly examined about the method of efficiently obtaining the proanthocyanidin containing containing high ratio of OPC which has useful physiological activity. As a result, it was found that by treating two or more kinds of adsorbents of plant extracts or juices, proanthocyanidin-containing compounds containing a large amount of physiologically active OPC were efficiently obtained, thereby completing the present invention.

The method for producing a proanthocyanidin-containing content of the present invention includes a step of treating an extract or juice of a plant with two or more adsorbents, each adsorbent having one or more of a range of material, micropore radius, specific surface area and molecular weight fraction. It differs in that point.

In a preferred embodiment, at least one of the adsorbents is a synthetic adsorbent.

In a preferred embodiment, the method is carried out using two kinds of adsorbents, wherein the first adsorbent is a synthetic adsorbent, the second adsorbent is a synthetic adsorbent, a cation exchange resin, an anion exchange resin, a crosslinked dextran derivative, polyvinyl It is selected from the group consisting of resins, agarose derivatives and cellulose derivatives.

In a preferred embodiment, one or more of the adsorbents can remove at least five protonocyanidins or contaminants from the extract or juice of the plant.

In a preferred embodiment, at least one of the adsorbents is porous and has a micropore radius of 90 kPa or less or 10 kPa or more.

In a preferred embodiment, at least one of the adsorbents has a molecular weight fraction range of 100 to 20,000 molecular weight.

Best Mode for Carrying Out the Invention

The method for producing a proanthocyanidin-containing product of the present invention includes a step of treating an extract or juice of a plant with two or more kinds of adsorbents, wherein the two or more kinds of adsorbents each have a material, specific surface area, micropore radius, and molecular weight fraction range. It differs in at least one of the points. The proanthocyanidins content obtained according to the preparation method of the present invention contains abundantly OPC, which is 2-4 tetramer of proanthocyanidins.

Extract or juice of plant

First, extracts or juices of plants are obtained.

The plant used in the present invention is not particularly limited as long as it is a plant containing proanthocyanidins. Tree bark of plants such as cedar, gingko and pine; Fruits, fruit peels and seeds of plants such as grapes, blueberries, strawberries, avocados, acacia, bilberry and elderberry; barley; wheat; Big head; Black soybeans; cacao; red bean; Fruit peel of horse chestnut; Thin shells of peanuts; Ginkgo biloba leaves; Tea leaves and tea extracts; Sorghum; Apple fruit; Stain stalks; Pcoidan; Yacon leaf; Cola nuts (eg, cola nuts in West Africa); Roots of latania (e.g., Latania, Peru). Among them, pine bark, grape seed and fruit bark, thin bark of peanuts and the like are suitably used.

When obtaining the extract of a plant, it carries out by adding an extraction solvent to a plant, and maintaining at predetermined temperature as needed.

In the case of extraction, in terms of extraction efficiency, the plant is preferably crushed to an appropriate size to increase the surface area per volume. The shredding method is not particularly limited. For example, the crushed object processed with the cutter, slicer, etc .; Pulverized products treated with a mixer, juicer, blender, mass collider, or the like. The size of the crushed or ground product is between 0.01 and 10 cm, preferably between 0.01 and 5 cm. In order to increase the crushing efficiency, water or organic solvents such as ethanol, methanol, ethyl acetate and the like can be added.

As the extraction solvent, water or an organic solvent is used. As an organic solvent, for example, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, acetone, hexane, cyclohexane, propylene glycol, hydrous ethanol, hydrous propylene glycol, ethyl methyl ketone, glycerin , Methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible fats and oils, 1,1,1,2-tetrafluoroethane and 1,1,2-trichloroethene. In addition, a mixed solvent of water-organic solvent is also preferably used. These organic solvents may be used alone or in combination. Moreover, water is used suitably from a viewpoint of the waste liquid process at the time of manufacture, or the point of adding the following metal or metal salt.

The amount of extraction solvent added to the plant can be set in consideration of the desired proanthocyanidin concentration and extraction efficiency. For example, when water is used as the extraction solvent, the ratio of plant and water is 1: 5 to 1: 100, preferably 1:10 to 1:50 in weight ratio. When crushing by adding water and / or an organic solvent, it is good to adjust the quantity of the extraction solvent to add in consideration of the quantity used for crushing.

The extraction temperature is preferably higher in order to increase the extraction efficiency. For example, when water is used, hydrothermal extraction is carried out at 50 to 120 ° C, preferably 70 to 100 ° C. Hot water may be added to the plant, and water may be added to the plant and then heated. The extraction time is generally 10 minutes to 48 hours, preferably 30 minutes to 24 hours, but may be appropriately determined depending on the extraction temperature. When the above heating is carried out, proanthocyanidins which are difficult to elute from plants can be efficiently extracted by being left again at 0 ° C to 30 ° C for 1 to 2 days. As an extraction method using an organic solvent, extraction by a warm extraction method and a supercritical fluid extraction method can be performed. As a warm extraction method, the method of adding the solvent heated to a plant, or the method of adding a solvent to a plant and warming is used. For example, using a water-ethanol mixed solvent having a ratio of water and ethanol of 1: 1 to 1: 9 in weight ratio to the ground plant, using 1 to 20 times the amount of the plant as an extraction solvent, while refluxing at 70 to 75 ℃ And a method of stirring for 0.5 hours to 6 hours. In the case of not refluxing, the mixed solvent is added once, then extracted by heating, the supernatant is recovered by filtration or the like, and the mixed solvent is added again to the remaining residue to increase the extraction efficiency. In addition, it is necessary to set the extraction temperature in the case of using an organic solvent below the boiling point of the said organic solvent.

The supercritical fluid extraction method is a method of extracting a target component using a supercritical fluid which is a fluid in a state of exceeding the critical point (critical temperature, critical pressure) of the gas liquid of a substance. As the supercritical fluid, carbon dioxide, ethylene, propane, nitrous oxide (gas) and the like are used, and carbon dioxide is preferably used.

The supercritical fluid extraction method consists of an extraction process of extracting a target component by a supercritical fluid and a separation process of separating the target component and the supercritical fluid. In the separation step, either extraction separation by pressure change, extraction separation by temperature change, or extraction separation using an adsorbent / absorbent can be performed.

In addition, supercritical fluid extraction can be performed by an entrainer addition method. This method is obtained by adding, for example, about 2 to 20 W / V% to a supercritical fluid by adding ethanol, propanol, n-hexane, acetone, toluene, other aliphatic lower alcohols, aliphatic hydrocarbons, aromatic hydrocarbons or ketones. The supercritical fluid extraction is performed with the extracted extraction fluid, which is a method of dramatically increasing the solubility in the extraction fluid of the target extract, such as OPC and catechins (hereinafter referred to), or enhancing the selectivity of separation. This is an efficient way to extract the din.

For extraction, any one extraction apparatus such as a batch, semi-continuous or continuous type can be used.

In the case of obtaining the juice of the plant, the plant is pressed directly, or after appropriately cutting or crushing, it is pressed. This method is suitably employed in the case of using plants having a high moisture content. For example, in the case of grape fruit, a juice containing proanthocyanidins is obtained by pressing. Plant shreds (such as grape fruit shreds) containing solids derived from plants obtained by shredding the plants are also used in the same manner as the juice, and plant shreds including solids derived from these plants are included in the juice in the present specification. .

Adsorbent

In the method of the present invention, two or more adsorbents are employed as described above. The individual adsorbents used in the present invention are not particularly limited as long as they are generally fillers used in column chromatography. Examples of the adsorbent include synthetic adsorbents, cation exchange resins, anion exchange resins, crosslinked dextran derivatives, polyvinyl resins (used as gels), agarose derivatives, cellulose derivatives, silica gels, reversed phase silica gels, and the like. Can be mentioned. Preferably, synthetic adsorbents, cation exchange resins, anion exchange resins, crosslinked dextran derivatives, polyvinyl resins (used in gel form), agarose derivatives, cellulose derivatives and the like can be given. More preferably, it is a synthetic adsorbent. Here, a synthetic adsorbent does not have functional groups, such as an ion exchanger, and is porous and has a fine continuous hole (micropore), for example, can adsorb | suck by van der Waals force.

Synthetic adsorbents are further classified according to their materials, and classified into synthetic adsorbents such as aromatic synthetic adsorbents, substituted aromatic synthetic adsorbents, and acrylic synthetic adsorbents. These synthetic adsorbents differ in degree of hydrophilicity and hydrophobicity depending on materials. In view of the stability of the synthetic adsorbent, the adsorption efficiency of proanthocyanidin (OPC), and the separation and fractionation ability, the aromatic adsorbent is preferably an aromatic synthetic adsorbent.

An aromatic synthetic adsorbent is an adsorbent consisting of porous resins such as crosslinked styrene resins. Commercially available adsorbents include, for example, DIION® HP-10, HP-20, HP-21, HP-30, HP-40 and HP-50 (above, Mitsubishi Chemical Corporation); Amberlite® XAD-4, XAD-16, XAD-1180 and XAD-2000 (above, Organo Corporation); And Sepha Biz® SP-825, SP-800, SP-850, and SP-875 (above, Mitsubishi Chemical Corporation).

Substituted aromatic synthetic adsorbents are adsorbents composed of a hydrophobic resin having a strong bromine atom bonded to an aromatic nucleus of an aromatic polymer. Examples of commercially available adsorbents include Sepha Beads (registered trademark) SP-205, SP-206, and SP-207 (above, Mitsubishi Chemical Corporation).

The acrylic synthetic adsorbent is an adsorbent composed of a hydrophilic resin having a methacrylic acid ester polymer or the like as a skeleton. Examples of commercially available adsorbents include, for example, Diion® HP1MG and HP2MG (above, Mitsubishi Chemical Corporation); And Amberlite (registered trademark) XAD-7 (manufactured by Organo Corporation).

Since the synthetic adsorbent is porous having fine continuous pores (micropores) as described above, the target solute present in the solution can be separated by the effect of molecular sieve. That is, when the synthetic adsorbent is brought into contact with the solution, small solute molecules penetrate and diffuse to the inside of the synthetic adsorbent through their micropores. On the other hand, molecules larger than the size of these micropores cannot diffuse into the synthetic adsorbent and are not adsorbed.

As an adsorbent consisting of a cation exchange resin, for example, Amberlite (registered trademark) CG-4000, CG-5000, CG-6000, CG-8000, IR-116, IR-118 which is a resin having a sulfonate group as a functional group , IR-120B, IR-122, IR-124, XT-1007, XT-1009, XT-1002 (above, organo corporation) etc. are mentioned.

As an adsorbent which consists of anion exchange resin, OPTIPORE-XUS 40285.00 which is a weakly basic anion exchange resin which has quaternary amine as a functional group, OPTIPORE-XUS 40390.00 (above, Dow Chemical Company) etc. are mentioned, for example.

Examples of the adsorbent composed of the crosslinked dextran derivatives include Sephadex® LH20 and LH60 (above, Amersham Biosciences Co., Ltd.).

Examples of the adsorbent made of polyvinyl resin (gel) include Toyo Pearl HW-40, 50 (Toso Corporation), and the like.

Examples of the adsorbent composed of agarose derivatives include Sepharose CL, 4B and 6B (above, Amersham Bioscience Company), and Bio-GelA (Biorad Laboratories Inc.).

As an adsorbent which consists of cellulose derivatives, cellophane CL-90, GCL-300, GCL-1000 (above, Seigagaku Corporation) etc. are mentioned.

Among the adsorbents described above, it is particularly preferable that the adsorbent be porous and have a networked molecular structure. For example, an adsorbent composed of a synthetic adsorbent such as Diion HP-20, Amberlite XAD-4, and an adsorbent composed of crosslinked dextran derivatives such as Sephadex LH20 and LH60 are particularly preferable.

The two or more adsorbents described above are different in one or more of each material, micropore radius, specific surface area, and molecular weight fraction range. For example, even if an adsorbent of the same material is used, an adsorbent having a different micropore radius, specific surface area or molecular weight fraction range is treated as a different kind of adsorbent. It is preferable that at least one type of adsorbent used is an adsorbent capable of removing pentaocyanate or more proanthocyanidins or an adsorbent capable of removing contaminants, and more preferably include both adsorbents.

The adsorbent capable of removing the proanthocyanidins of the pentamer or more (hereinafter referred to as the first adsorbent) is specifically capable of adsorbing OPC and adsorbing the proanthocyanidins of the pentamer or more. It is a difficult adsorbent. An adsorbent capable of removing a contaminant (hereinafter referred to as a second adsorbent) is specifically an adsorbent which easily adsorbs proanthocyanidins and which is difficult to adsorb a contaminant, preferably in addition to these properties Furthermore, after adsorbing proanthocyanidins, OPC is selectively separated and fractionated from the proanthocyanidins and eluted to obtain a solution having a high OPC content.

In the case where the first adsorbent and the second adsorbent are selected as the material standards, the aromatic synthetic adsorbent is preferably selected as the first adsorbent. The second adsorbent is preferably selected from the group consisting of synthetic adsorbents, cation exchange resins, anion exchange resins, crosslinked dextran derivatives, polyvinyl resins, agarose derivatives and cellulose derivatives. More preferably a synthetic adsorbent or crosslinked dextran derivative is selected, particularly preferably an aromatic synthetic adsorbent or crosslinked dextran derivative.

As the adsorbent, a porous synthetic adsorbent having micropores as described above is preferable, and the first adsorbent and the second adsorbent can be selected based on the micropore radius. This is because the size of the molecules to be adsorbed or the adsorption force to the molecules is different depending on the micropore radius.

The first adsorbent preferably has a micropore radius of 90 kPa or less, more preferably 20 to 90 kPa, still more preferably 30 kPa to 80 kPa. The smaller the fine pore radius, the higher the adsorption capacity for low molecules (thousands of molecular weight or less) than polymers. Therefore, while adsorb | sucking a low molecular weight compound, it can remove without adsorb | sucking the proanthocyanidin more than pentamer which is comparatively high molecular weight. As such an adsorbent, for example, Sephabiz SP-825, Sephabiz SP-850, Amberlite XAD-4, and XAD-2000 which are adsorbents composed of an aromatic synthetic adsorbent are suitable.

The second adsorbent preferably has a fine pore radius of 100 kPa or more, more preferably 100 kPa to 500 kPa, still more preferably 100 kPa to 300 kPa. The larger the fine pore radius, the more efficiently the proanthocyanidins having a wide range of molecular weights ranging from tens of thousands to tens of thousands of molecules are used. However, since the force of adsorbing the contaminants is weak, it can be easily removed without adsorbing the contaminants. As such an adsorbent, Sephadex LH20, which is an adsorbent composed of diion HP-20, diion HP-21 and amberlite XAD-16 or a crosslinked dextran derivative, is an adsorbent composed of an aromatic synthetic adsorbent, for example. .

In the case where the first adsorbent and the second adsorbent are selected on the basis of the specific surface area, it is preferable that both of the first adsorbent and the second adsorbent have a specific surface area of 500 m ² / g or more. In addition, from the viewpoint of efficiently adsorbing OPC, it is more preferable that the specific surface area is 70Om ² / g or more as the first adsorbent. The specific surface area of the adsorbents varies depending on the size of the individual adsorbents, or in the case of porous adsorbents, depending on the size and number of the micropores, so that an adsorbent of the required specific surface area can be appropriately selected.

In the case of fractionation according to molecular weight using an adsorbent such as a crosslinked dextran derivative or a polyvinyl resin (which uses a gel phase) as the adsorbent, the molecular weight fraction range is not particularly limited. It is preferred that the fraction range is 100 to 250,000, preferably 100 to 5,000. The second adsorbent preferably has a molecular weight fraction in the range of 100 to 20,000, preferably 100 to 10,000. Adsorbents having this molecular weight fraction range can adsorb proanthocyanidins and remove contaminants. Furthermore, OPC in adsorbed proanthocyanidins can be eluted and fractionated. Of these, Sephadex LH-20 and Sephadex LH-60 are suitable.

Among the various adsorbents, as the first adsorbent, Amberlite (registered trademark) XAD-4 and Sepha Beads SP825, which are porous, have an adsorbent composed of an aromatic synthetic adsorbent having a specific pore radius of 90 mm or less and a specific surface area of 700 m ² / g or more. And SP850 are particularly suitable. As the second adsorbent, it is an adsorbent composed of a diaion® HP-20 and a crosslinked dextran derivative, which is an adsorbent made of an aromatic synthetic adsorbent having a pore diameter of 100 microns or more and a specific surface area of 500 m ² / g or more. Sephadex LH20 is particularly suitable.

The amount of the adsorbent may be appropriately set depending on the amount of solids contained in the object to be treated (extract or juice of the plant or the recovered matter from the adsorbent), the kind of the solvent (described below), the kind of the adsorbent, and the like. For example, it is preferable to use 0.1-100 weight part, Preferably 0.1-50 weight part adsorption body with respect to 1 weight part of dry weights of solid content contained in a to-be-processed object. If it is less than 0.01 part by weight, the recovery rate of proanthocyanidin is lowered. If it is more than 100 parts by weight, it can be sufficiently adsorbed. However, the recovery from the adsorbent is poor, which is not preferable. In the case of using an adsorbent having a high adsorption efficiency of OPC, the amount of the adsorbent can be set based on the weight of the plant before the treatment without measuring the dry weight in the workpiece for simpler operation. For example, when using an adsorbent made of an aromatic synthetic adsorbent such as DIION HP-20, Sephabiz SP-825, Amberlite XAD-16, or an adsorbent made of crosslinked dextran derivatives such as Sephadex LH20 For example, the adsorbent is used in an amount of 0.1 to 10 parts by weight, preferably 0.2 to 5 parts by weight, based on 1 part by weight of the dry weight of the plant before treatment, so that the contact between the object and the adsorbent is sufficient. And can be adsorbed efficiently. Specifically, when the pine bark is treated with Diion HP-20, 0.1 kg to 10 kg (equivalent to 0.15 L to 15 L of the adsorbent body) is used for 1 kg of pine bark. In the case of using an adsorbent of crosslinked dextran derivatives, the volume is swollen and the volume is increased when water is added. Therefore, the volume is swelled in a volume of 100 to 1000 volumes based on 1 part by weight of the dry weight of the solid content contained in the treatment liquid. Preferably, it is preferable to use an adsorbent of 120 to 500 volume parts.

Adsorbent treatment

According to the method of the present invention, a proanthocyanidin-containing product is treated by treating extracts or juices of plants using a combination of two or more kinds of adsorbents described above, followed by treatment such as concentration treatment and salt treatment, if necessary. Obtained. For example, by contacting the extract or juice of the plant with the first adsorbent, at least pentameric proanthocyanidins that are not adsorbed are removed, and the adsorbate is recovered with a predetermined solvent, and then using another adsorbent, From the recovered product can be obtained proanthocyanidins containing a high proportion of OPC. In this case, the order of the treatment by the first adsorbent and the treatment by the other adsorbent may be reversed. Alternatively, the extract or juice of the plant is contacted with a second adsorbent to remove contaminants as nonadsorbents, and then another adsorbent is used to obtain a proanthocyanidin content containing a high proportion of OPC from the recovered product. can do. In this case, the order of the treatment by the second adsorbent and the treatment by the other adsorbent may be reversed. It is more appropriate to carry out combining the treatment by the first adsorbent and the treatment by the second adsorbent. In this case, the order of the treatment by the first adsorbent and the treatment by the second adsorbent may be reversed. Moreover, it can process with three or more types of adsorbents by performing the process by another adsorbent suitably.

Contact of the object (extract of plant or extract from juice or adsorbent) with the adsorbent may be carried out by any method. For example, as a simple method, a column method in which an adsorbent is filled in a column, the column is passed through the column, and an adsorbent is added to the object, and a batch method of removing the adsorbent after a certain time is mentioned. Can be.

In the treatment using the column method, first, if necessary, the solvent of the workpiece is replaced with a solvent suitable for contact with the adsorbent. Such substitution may be carried out according to a method including a process commonly used by those skilled in the art, such as heat drying, freeze drying, reduced pressure drying, dialysis, and the like. For example, in the case of an adsorbent made of an aromatic synthetic adsorbent such as diion HP-20, it is substituted with water, and in the case of an adsorbent made of a crosslinked dextran derivative such as Sephadex LH20, it is substituted with ethanol. The second adsorbent is then charged to the column, and the extract or juice of the plant or the recovered product is passed through the column, and, for example, 3 to 10 times the volume of water is passed through the volume of the adsorbent. . As a result, the sugars and organic acids that are contaminants are removed. Subsequently, the proanthocyanidins are eluted with an appropriate solvent (described below). In addition, what is necessary is just to determine suitably the various conditions in a column method according to the adsorption body used, For example, when using the adsorption body which consists of ion-exchange resin, column temperature is set to 10 to 120 degreeC, It is preferable to make the inside of a column normal or pressurized.

In the treatment using the batch method, an adsorbent in the same weight ratio as the column method is added to the to-be-processed object, contacted for 1 to 3 hours with stirring, and then the adsorbent is recovered by filtration or centrifugation. For example, when using a 2nd adsorption body as an adsorption body, a contaminant can be removed by such an operation. Subsequently, the second adsorbent adsorbed with proanthocyanidin is again stirred with an appropriate solvent (below) for 1 to 3 hours, the proanthocyanidin is eluted, and then filtered or centrifuged to recover the supernatant. Proanthocyanidins or higher containing proanthocyanidins can be obtained.

The eluting solvent may be appropriately selected depending on the kind of the adsorbent and the kind of the substance to be adsorbed or eluted. For example, water, methanol, ethanol, ethyl acetate, chloroform, and these mixed solvents are mentioned as an elution solvent in the case of using a 1st adsorption body. In terms of safety, preferably a mixed solvent of water and ethanol is used. The mixing ratio of water and ethanol is, for example, an adsorption made of an aromatic synthetic adsorbent which removes proanthocyanidins of at least 5 mers, such as Amberlite XAD-4, Amberlite XAD-2000, Sephabiz SP825, Sephabiz SP850, etc. In the case of a sieve, it is adsorbed to use a relatively high concentration of an aqueous ethanol solution of at least 10% by volume, preferably at least 30% by volume, more preferably at least 50% by volume, in that at least 5mers of proanthocyanidins are removed. It is preferable from the viewpoint of increasing the recovery rate when eluting the OPC.

As the elution solvent in the case of using the second adsorbent, proanthocyanidins are adsorbed regardless of the molecular weight, so it is preferable to select a solvent capable of selectively eluting OPC from the adsorbed proanthocyanidins to separate and fractionate it. Do. Preferably, it is a mixed solvent (ethanol aqueous solution) of water and ethanol. For example, in the case of an adsorbent composed of aromatic synthetic adsorbents such as diion HP-20, HP-21, XAD-16, a proanthocyanidin-containing substance containing a large amount of OPC is separated from the adsorbate. For this purpose, an aqueous ethanol solution of 10% by volume to 50% by volume, preferably 10% by volume to 30% by volume is preferred. In the case of an adsorbent composed of crosslinked dextran derivatives such as Sephadex LH-20 and Sephadex LH-60, an aqueous ethanol solution of 70% by volume or more, preferably 80% by volume or more is preferable.

When using the adsorbent which consists of ion exchange resin, it is preferable to use water as an elution solvent.

By using different kinds of adsorbents as described above, it is possible to increase the recovery rate of 2-4 tetraproanthocyanidins (OPC) from extracts or juices of plants, and OPC content in the obtained proanthocyanidins content Can be increased efficiently.

Concentrate and salt treatment

As described above, the concentration treatment or the salt treatment can be carried out as necessary, before or after the two or more adsorbent treatments or between one adsorbent treatment and the next adsorbent treatment.

Concentration treatment increases the adsorption efficiency to the adsorbent and at the same time, the proanthocyanidins of pentamers or more may form precipitates by concentration, so that the proanthocyanidins having a higher content of OPC are removed by filtration or the like. Dean content can be obtained. In the case of adsorption treatment using an adsorbent made of an aromatic synthetic adsorbent, the solvent needs to be water due to the nature of the resin. Therefore, if an organic solvent is contained in the extract or juice of the plant, it is concentrated before the adsorbent treatment. The removal of the solvent can also be carried out simultaneously.

The concentration treatment may be carried out according to a concentration method commonly used by those skilled in the art, such as vacuum concentration and freeze drying. In the case of concentration by heating, in order to prevent denaturation of proanthocyanidin by heating, it is carried out at a temperature of 40 ° C to 100 ° C. A reduced pressure concentration method or a freeze drying method with less modification of proanthocyanidins is preferable.

In addition, in order to perform a concentration process at a relatively low temperature for a short time, it is preferable that the solution used for a to-be-processed object is an organic solvent, such as ethanol, whose boiling point is lower than water, or a mixed solution of these organic solvents and water. Ethanol or a mixed solution of ethanol and water is also suitable from the viewpoint of safety when used as food or medicine.

Concentration treatment is preferably carried out after filtration once to remove the insoluble matter of the to-be-processed object, so that the degree of concentration may be easily produced.

There is no particular limitation on the concentration rate of the obtained concentrate. The volume of the concentrate is preferably from 1/2 to 1/100 volume, more preferably from 1/5 to 1/70 volume, more preferably from 1/10 to l / 50, compared to the volume of the workpiece before concentration. Concentration is carried out to ensure volume.

Salt treatment can remove such insoluble matters by adding a salt to a to-be-processed object, and generate | occur | producing a high degree of polymerization proanthocyanidins, for example as insolubles, such as a precipitate. In the salt treatment, it is preferable to replace the solvent in the object to be treated with water. Substitution with water improves the ionization of the salt solution and can efficiently remove proanthocyanidins having a high degree of polymerization.

There is no particular limitation on the temperature of the workpiece when adding the salt. It is preferable to add a salt after falling to 1-40 degreeC.

The salt added to the to-be-processed object should just be ionized in solution. As a salt, a monovalent metal salt, a bivalent metal salt, and a nonmetallic salt are mentioned, for example.

As monovalent metal salt, salts, such as lithium, sodium, potassium, rubidium, cesium, and francium which are alkali metals, are suitable.

Examples of the divalent metal salts include metal salts such as beryllium, magnesium, and alkaline earth metals (calcium, strontium, barium, and radium).

As a nonmetallic salt, ammonium sulfate etc. are used suitably, for example.

Since the divalent metal salt has a particularly strong adsorptive force to proanthocyanidins, the divalent metal salt is bound not only to proanthocyanidins having a high degree of polymerization but also to OPC, and is likely to cause precipitation. In addition, when a divalent metal salt is added, since the pH in a solution tends to become high, there exists a possibility that a proanthocyanidin may be oxidized. The divalent metal salt is added so that the pH in the solution is preferably less than 7.5, more preferably less than 6, still more preferably 5.5 or less. When pH becomes 7.5 or more, it is good to add the adjuvant (for example, pH adjusters, such as ascorbic acid) for stabilizing proanthocyanidin beforehand. In addition, divalent metal salts require caution because proanthocyanidins may be oxidized in the same manner as described above when a salt containing a metal (for example, copper) used as an oxidizing agent is used. Thus, monovalent alkali metal salt or ammonium sulfate is preferable at the point which can make OPC and a salt as small as possible and improve OPC refinement | purification efficiency.

Examples of such monovalent metal salts include halide salts (chloride salts, bromide salts, etc.), phosphates, carbonates, organic acid salts (carboxylates such as acetates, sulfonates, and the like). Specifically, sodium chloride, sodium sulfate, sodium citrate, potassium chloride, sodium phosphate, potassium phosphate, sodium acetate, etc. are mentioned. Particularly preferred are sodium sulfate, potassium phosphate, sodium citrate, sodium chloride and ammonium sulfate, which are preferably used for salting out.

The salt is different depending on the type of salt, but may be added so as to be 3 to 50% by weight, preferably 5 to 45% by weight, based on the total weight of the object to be treated. Further, when the maximum dissolving amount of the salt in water is preferably 100 parts by weight, an amount corresponding to 10 to 75 parts by weight, more preferably 20 to 60 parts by weight is added to the concentrate. In addition, when using divalent metal salts, such as a calcium salt and magnesium salt, as a salt, it is good to add so that it may become 0.1 to 30 weight% in a concentrate.

When treating with a salt, in particular with a metal salt, treatment on the acidic side is preferred. In weak alkaline to strong alkali, the stability of proanthocyanidins deteriorates and there is a possibility of decomposition. Therefore, in the case of using such a metal salt, after adjusting the pH of the solution at a concentration of 2 to 10 times the predetermined final concentration to 4 to 6, preferably 4 to 5.5, more preferably 4 to 5, such a solution It is preferable to add to the to-be-processed object.

After the salt is added, it is allowed to stand for 30 minutes to 48 hours at, for example, 1 ° C. to 40 ° C., and sufficiently generates an insoluble matter such as a precipitate. In addition, the settling time may be 48 hours or more, but it is preferable to move to the next step before the OPC automatically oxidizes and the reddish brown turns to dark brown.

Subsequently, insoluble matters, such as a precipitate which generate | occur | produced, are removed. As a method of removing an insoluble matter, the method normally used by a person skilled in the art, for example, filtration, centrifugation, etc. are mentioned. In terms of treatment time, filtration is suitably used. Filtration may preferably be carried out at 1 to 40 ° C. The lower the temperature, the more proanthocyanidins having a higher degree of polymerization can be removed, and preferably at 30 ° C or lower, more preferably at 25 ° C or lower. Such filtration can be carried out before the addition of salt, but it is necessary to filter to remove insoluble matters such as precipitates even after addition of salt. In addition, in order to minimize the loss by filtration, the residue remaining after the filtration can be washed with an aqueous solution having the same saturation concentration, and the washing liquid can be recovered.

In the present invention, when the salt treatment is performed before the treatment in the adsorbent, the salt may be removed in the step of treating with the adsorbent, so that the salt removal step is not necessarily required.

Proanthocyanidins Contain

The proanthocyanidins content thus obtained contains a high proportion of proanthocyanidins. Proanthocyanidins are proanthocyanidins having a low degree of polymerization in terms of physiological activity, preferably condensates having a degree of polymerization of 2 to 30 (2 to 30 monomers), more preferably condensates having a degree of polymerization of 2 to 10. (2 to 10 monomers), more preferably a condensate (2 to 4 monomers; OPC) having a degree of polymerization of 2 to 4 is preferred. The proanthocyanidins content obtained according to the process of the invention contains especially high proportions of OPC, preferably at least 35% by weight, more preferably at least 40% by weight, in terms of dry weight. In addition, since the ratio of proanthocyanidins of pentamer or more is 1 part by weight or less with respect to 1 part by weight of OPC, solubility and physiological activity in water is high, and excellent proanthocyanidins with reduced astringent taste and the like It becomes an artifact.

The proportion of OPC in the total proanthocyanidins contained in the proanthocyanidin content obtained according to the present invention is preferably at least 45% by weight, more preferably at least 50% by weight. As such, the proanthocyanidins content obtained by treatment with two or more adsorbents is compared to the extract or juice of the plant, or the treatment obtained by treatment with a single adsorbent, to the OPC fraction of the total proanthocyanidins. The ratio is high and preferably 3 times or more, preferably 3.5 times or more, as compared with the extract or juice of the plant.

The proanthocyanidin content obtained according to the method of the present invention may further contain catechins, preferably in the range of dry weight, in the range of 10 to 15% by weight. Although catechins alone lack water solubility and their physiological activity is low, OPC, when coexisted with catechins, increases the water solubility of catechins and at the same time activates the action of catechins. Therefore, proanthocyanidins containing OPC and catechins are particularly useful.

Catechins are a general term for polyhydroxyflavan-3-ol. Examples of catechins include (+)-catechin, (-)-epicatechin, (+)-gallocatechin, (-)-epigallocatechin, epigallocatechin gallate, epicatechin gallate and the like. Also included are azelechins derived from natural products and 3-galloyl derivatives of (+)-catechin or gallocatechin.

The effects of catechins include carcinogenesis inhibitory action, arteriosclerosis prevention action, fat metabolism inhibition action, blood pressure increase suppression action, platelet aggregation inhibition action, antiallergic action, antiviral action, antibacterial action, tooth decay prevention, bad breath prevention action, intestinal tract Bacterium normalizing action, scavenging action of free radicals and free radicals, antioxidant action, antidiabetic action that inhibits blood sugar rise.

Subsequently, the proanthocyanidin-containing content having a very high OPC content according to the present invention can be made into concentrates, diluents and the like according to methods commonly used by those skilled in the art. Various methods, such as membrane concentration, heat concentration, vacuum (pressure reduction) concentration, and freeze concentration, are used for concentration. If necessary, these proanthocyanidin-containing contents are sterilized and stored. Sterilization is carried out in accordance with methods commonly used by those skilled in the art, such as air sterilization, autoclaving and heat sterilization.

In addition, these proanthocyanidins contents can be concentrated, dried and powdered after sterilization. Drying is carried out in accordance with methods commonly used by those skilled in the art. Among these, freeze drying, vacuum drying and spray drying are preferably used.

The proanthocyanidin-containing materials thus obtained can be used as raw materials for food, medicine, cosmetics and quasi-drugs.

When using the proanthocyanidin content obtained by this invention as a foodstuff, it is good also as a drink agent, the gelatinized foodstuff, etc. In addition, the proanthocyanidins content may be provided to the food as is or by adding an additive. Excipients, extenders, binders, thickeners, emulsifiers, flavorings, nutritional ingredients, food additives, seasonings and the like can be given as additives. For example, royal jelly, vitamins, protein, calcium, chitosan, lecithin, caffeine, and the like are blended as nutritional ingredients, and as a food additive, sugar and seasonings can be added to provide flavor. These may be shaped into capsules, tablets or pills, such as hard capsules and soft capsules, or powders, granules, syrups, or the like as necessary. And they can be eaten as it is according to the shape or preference thereof, and can be rinsed with water, hot water, milk or the like, or put into a tea bag or the like, and drinkable with a bath or the like.

Even when the proanthocyanidin-containing compound obtained according to the present invention is used as a medicine, it can be molded into the same shape as that of the food, as it is or using additives commonly used by those skilled in the art.

When the proanthocyanidin content obtained according to the present invention is used as cosmetics and quasi-drugs, the solubility of aqueous solutions is excellent, so that those skilled in the art such as ointments, creams, emulsions, lotions, lotions, packs and bath preparations are commonly used. It can be molded into a shape to be.

Hereinafter, although this invention is demonstrated based on an Example, this Example does not limit this invention. The unit (V / V) described in the Example represents (volume / volume), and (W / W) represents (weight / weight).

Preparation of Extract

5.4 L of purified water was added to 1 kg of pine bark, crushed with a Waring Blender, and extracted by heating under reflux at 100 ° C. for 24 hours. Subsequently, the filtrate was immediately filtered, the insoluble matters after filtration were washed with 1.6 L of purified water, and the filtrate and the washing liquid were combined to obtain 7 L of pine bark extract. 10 mL of this extract was lyophilized and the dry weight was 70 mg. This extract is allowed to cool to 25 ° C.

Example 1

Amberlite (registered trademark) XAD-4 [Organocorporation agent] comprising 1 L (extract powder dry weight 7 g) of the extract, an aromatic synthetic adsorbent having a specific surface area of 70Om ² / g and a micropore radius of 80 kPa or less; Adsorbent (first adsorbent) capable of removing proanthocyanidins of 5 or more] is passed through a column of 5 cm diameter filled with 300 mL (approximately 200 g). This column is then washed with 600 mL of purified water. In addition, 800 mL of 80 V / V% aqueous ethanol solution was passed through to elute the adsorbate. This treated solution was concentrated under reduced pressure, ethanol was removed, and then adjusted to 500 mL by adding water. Subsequently, such a solution was prepared by diion (registered trademark) HP-20 [Mitsubishi Chemical Corporation] consisting of an aromatic synthetic adsorbent having a specific surface area of 600 m ² / g and a fine pore radius of 100 to 120 kPa; Adsorbent (second adsorbent) capable of removing contaminants] is passed through a column of 5 cm diameter filled with 200 mL (approximately 140 g equivalent). After washing this column with 600 mL of purified water, 200 mL of 20 V / V% aqueous ethanol solution was passed through, and 500 mL was passed through again to combine them. Let the obtained solution be a proanthocyanidin containing liquid (A).

Subsequently, in order to examine the content of each component in the obtained proanthocyanidin-containing liquid (A), the component in the proanthocyanidin-containing liquid (A) was separated into an OPC fraction, a pentameric or higher proanthocyanidin fraction, A fraction containing catechins and a fraction of components other than catechins are separated as follows. The proanthocyanidin-containing liquid (A) is concentrated to dryness under reduced pressure, to a dry powder, and the weight thereof (solid weight) is measured. 25 mL of Sephadex LH-20 (made by Amersham Biotech Co., Ltd.) swollen with water is charged into a 15 x 300 mm column and washed with 50 mL of ethanol. 100 mg of the dry powder is dissolved in 2 mL of ethanol, the solution thereof is passed through a column to be adsorbed, gradient eluted with a 100 to 80 V / V% ethanol-water mixed solvent, and aliquots are separated by 10 mL. Standards of 2-4 tetrameric OPC [Dimer: proanthocyanidin B-2 (Rf value: 0.6), trimer: proanthocyanidin C-1 (Rf value: 0.4) and tetramer when fractionated : Cinnamic tannin A2 (Rf value: 0.2)] is used as an index to detect the presence or absence of OPC in each fraction by silica gel thin layer chromatography (TLC) under the following conditions.

TLC: silica gel plate (manufactured by Merck & C0., Inc.)

Developing Solvent: Benzene / Ethyl Formate / Formic Acid (2/7/1)

Detection reagents: sulfuric acid and anisealdehyde sulfuric acid

Sample volume: 10 μL each

The elution fractions confirming that OPC is contained by TLC are combined to obtain an OPC fraction.

Then, at the point where OPC was not detected, 300 mL of 50% (V / V) water-acetone mixed solvent was passed through, and the remaining adsorbate adsorbed on the column was eluted.

Elution fractions other than the recovered OPC fractions were subjected to TLC to separate fractions containing catechins and fractions containing at least 5 grams of proanthocyanidins. The development conditions and the detection method of TLC are performed similarly to the above.

In addition, the fraction containing catechins is separated into components other than catechins and catechins as follows. First, the fraction containing catechins is lyophilized and a powder is obtained. This powder is dissolved in 3 mL of water, and 20 mL of MCI gel (manufactured by Mitsubishi Chemical Corporation) in which this solution is swollen with water is passed through a column packed into a 15 x 300 mm column and adsorbed. This column is washed with water, then eluted with 10 to 100% (V / V) ethanol-water mixed solvent and aliquoted 7 mL. After the end of the elution, catechins in each fraction are detected by TLC using catechin as an index and divided into catechin fractions and fractions other than catechins.

OPC fractions obtained as described above, pentameric or higher proanthocyanidin fractions, catechin fractions and fractions of components other than catechins were respectively powdered by freeze drying and the dry weight was measured. In addition, the sum total of OPC fraction, pentameric or more proanthocyanidin fraction, catechin fraction, fraction of components other than catechins, and the fraction of other components was made with respect to 100 mg of dry powder of a proanthocyanidin containing liquid (A). It was found that it was 99.0 to 99.9 mg, and almost the entire amount was recovered.

Table 1 shows the solids weight of the proanthocyanidin-containing solution (A), OPC contained in the proanthocyanidin-containing solution (A), proanthocyanidins of 5 or more weights, total proanthocyanidins (OPC and The total weight of proanthocyanidins of pentamer or more) and the dry weight and content of catechins, and the ratio of OPC in total proanthocyanidins. Table 1 further describes the treatment conditions of the adsorbent. (Circle) mark shows using a 1st or 2nd adsorption body, and x mark shows not using a 1st or 2nd adsorption body.

Example 2

The same procedure as in Example 1 was conducted except that the order of treatment with the first adsorbent and the treatment with the second adsorbent were reversed to obtain a proanthocyanidin-containing liquid (B), which was the same as in Example 1. Analyze The results are shown in Table 1.

Comparative Example 1

The treatment liquid obtained by treating in the same manner as in Example 1 was analyzed in the same manner as in Example 1 except that the treatment with the second adsorbent was not carried out. The results are shown in Table 1.

Comparative Example 2

Treatment was carried out in the same manner as in Example 1 except that no treatment with the second adsorbent and an aqueous 20V / V% aqueous ethanol solution were used instead of the aqueous 80V / V% aqueous solution of the treatment with the first adsorbent. The treated solution is analyzed in the same manner as in Example 1. The results are shown in Table 1.

Comparative Example 3

A treatment liquid obtained by treating in the same manner as in Example 1 was analyzed in the same manner as in Example 1 except that the treatment with the first adsorbent was not carried out. The results are shown in Table 1.

Comparative Example 4

Treatment was carried out in the same manner as in Example 1 except that no treatment with the second adsorbent and an aqueous 80V / V% aqueous ethanol solution were used instead of the 20V / V% aqueous ethanol solution for the treatment with the second adsorbent. The treated solution is analyzed in the same manner as in Example 1. The results are shown in Table 1.

Comparative Example 5

The same treatment as in Example 1 was carried out except that 5V / V% aqueous ethanol solution was used instead of 20V / V% aqueous ethanol solution without treatment with the first adsorbent and with the second adsorbent. The treatment liquid is analyzed in the same manner as in Example 1. The results are shown in Table 1.

Comparative Example 6

The treatment liquid obtained by treating in the same manner as in Example 1 was analyzed in the same manner as in Example 1 except that the treatment with the first adsorber was again performed instead of the treatment with the second adsorber. The results are shown in Table 1.

Comparative Example 7

The treatment liquid obtained by treating in the same manner as in Example 1 was analyzed in the same manner as in Example 1 except that the treatment by the second absorbent was again performed instead of the treatment by the first absorbent. The results are shown in Table 1.

Comparative Example 8

The extract from pine bark is analyzed in the same manner as in Example 1 instead of the proanthocyanidin-containing solution (A). The results are shown in Table 1.

From the results in Table 1, it was found that in Examples 1 and 2 using two different kinds of adsorbents, proanthocyanidin-containing compounds containing a high proportion of OPC were obtained. In particular, Examples 1 and 2 combine an adsorbent (first adsorbent) that elutes and removes proanthocyanidins of pentamer or more, and an adsorbent (second adsorbent) that removes contaminants and fractionates OPC. It was found that OPC was contained by 40% by weight or more in the resulting dry powder.

The adsorbents used in Comparative Examples 1 and 4 differ in particular in terms of micropore radius. In Comparative Example 1 using an adsorbent having a micropore radius of 90 kPa or less, the OPC content is higher than that of Comparative Example 4 using an adsorbent having a micropore radius of 100 kPa or more. On the other hand, Comparative Example 4 has a higher total proanthocyanidin content in the dry powder than in Comparative Example 1. In this way, when the adsorbent having a micropore radius of 90 kPa or less is used, when proanthocyanidin is adsorbed to the adsorbent, proanthocyanidins of 5 or more are easily removed, so that the OPC content of the treated product is relatively high. If an adsorbent having a micropore radius of 100 kPa or more is used, the total proanthocyanidin content of the treated product tends to be relatively high. Comparing Comparative Examples 1 and 2, in the case of using an adsorbent having a micropore radius of 90 kPa or less, even if the extraction solvent was changed, the OPC content did not change. Compared to Comparative Examples 3 and 4, the micropore radius was 100 kPa. In the case of using the above-described adsorbent, it was found that the OPC content was changed by changing the extraction solvent. From this point of view, in the case of selectively eluting OPC by the concentration of the solvent, it is considered that an adsorbent having a fine pore radius of 100 kPa or more is suitable. In Comparative Example 5, only one type of second adsorbent is used to obtain a proanthocyanidin-containing substance having a high OPC content by separating OPC and pentacyano dimer or more. In Comparative Example 5, an elution solvent (5 V / V% ethanol aqueous solution) under conditions of eluting only OPC was used, but it was found that OPC was not sufficiently eluted and the recovery amount of OPC itself was low.

Example 3

2 L of purified water is added to 100 g of pine bark, crushed with a Waring Blender, and heated at 95 ° C. for 1 hour. Then, the filtrate was immediately filtered, and the insoluble matter after filtration was washed with 1 L of purified water, and the filtrate and the washing liquid were combined to give 3 L of extract.

Subsequently, 70 g (equivalent to about 100 mL) of Sephabiz SP850 was added to 3 L of this extract, stirred for 3 hours, and filtered to recover the solid adsorbed with proanthocyanidins. This solid was washed with 250 mL of purified water, 150 mL of 80 (V / V)% ethanol aqueous solution was added, stirred for 1 hour, and then filtered to recover the filtrate. The filtrate is concentrated under reduced pressure to remove ethanol, and purified water is added to make 1 L. 70 g of Diaion® HP-20 was added to this solution, stirred for 3 hours, and then filtered to recover a solid to which proanthocyanidins were adsorbed, and the solid was washed with 250 mL of purified water. 150 mL of 15 (V / V)% ethanol aqueous solution was added, stirred for 1 hour, and filtered to recover the filtrate. This filtrate is lyophilized to yield 351 mg of proanthocyanidin containing powder. Using 100 mg of proanthocyanidin-containing dry powder, the content of OPC and catechins was measured in the same manner as in Example 1, whereby OPC 42.3% by weight in terms of dry weight of proanthocyanidin 10 or more Wt% and catechins 14.9 wt%.

Example 4

1 L of purified water is added to 100 g of pine bark, crushed with a Waring Blender, and heated at 100 ° C. for 12 hours. Then, the filtrate was immediately filtered, the insoluble matters after filtration were washed with 200 mL of purified water, and the filtrate and the washing liquid were combined to obtain 1.2 L of an extract. This extract is passed through a column filled with 80 g (equivalent to about 100 mL) of Amberlite® XAD-4. This column was washed with 300 mL of purified water and then passed through 600 mL of 20 (V / V)% ethanol aqueous solution to obtain an eluate. The eluate was concentrated under reduced pressure to dryness, and the powder obtained was dissolved in 500 mL of absolute ethanol again. The column packed with 300 mL of Sephadex LH-20 swelled with water was washed with 300 mL of ethanol, and the eluate was passed through. This column was washed with 600 mL of anhydrous ethanol and then passed through 1.5 L of 80 V / V% aqueous ethanol solution to obtain a proanthocyanidin-containing solution. The contents were concentrated under reduced pressure to dryness to yield 421 mg of proanthocyanidins. The content of OPC and catechins was measured in the same manner as in Example 1, and found to be 46.2% by weight of OPC, 55.2 or more, 15.2% by weight, and 11.1% by weight of catechins in terms of dry weight.

According to the method of the present invention, by treating the extract or juice of the plant with at least two kinds of adsorbents different in one or more points of material, specific surface area, micropore radius and molecular weight fraction range, it contains a high ratio of high physiologically active OPC. Proanthocyanidins content can be readily obtained. These proanthocyanidins are effective for improving blood vessel enhancement, hypertension, coldness, etc., and are very useful as raw materials for the preparation of food, medicine, cosmetics and quasi-drugs.

Claims (6)

In the manufacturing method of the proanthocyanidin-containing thing containing the process of processing extract or juice of a plant with two or more adsorbents, Wherein said adsorbents are each different in at least one of material, micropore radius, specific surface area and molecular weight fraction range. The method according to claim 1, wherein at least one kind in the adsorbent is a synthetic adsorbent. The method according to claim 1 or 2, wherein the first adsorbent is a synthetic adsorbent, the second adsorbent is a synthetic adsorbent, a cation exchange resin, an anion exchange resin, a crosslinked dextran derivative, Polyvinyl resin, agarose derivatives and cellulose derivatives. The method according to claim 1 or 2, wherein at least one kind of adsorbent can remove pentameric or more proanthocyanidins or contaminants from extracts or juices of plants. The method according to any one of claims 1 to 3, wherein at least one of the adsorbents is porous and has a micropore radius of 90 kPa or less or 100 kPa or more. The method according to any one of claims 1 to 4, wherein at least one of the adsorbents has a molecular weight fraction range of 100 to 20,000 in molecular weight.