WO2009099222A1 - Process and apparatus for production of composition having high polyphenol content - Google Patents
Process and apparatus for production of composition having high polyphenol content Download PDFInfo
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- WO2009099222A1 WO2009099222A1 PCT/JP2009/052118 JP2009052118W WO2009099222A1 WO 2009099222 A1 WO2009099222 A1 WO 2009099222A1 JP 2009052118 W JP2009052118 W JP 2009052118W WO 2009099222 A1 WO2009099222 A1 WO 2009099222A1
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- polyphenol
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- eluate
- exchange resin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Definitions
- the present invention relates to a polyphenol-rich composition and a method for producing the same. More specifically, the present invention relates to a method for efficiently producing a composition having a low theobromine content and a high polyphenol content by selectively extracting the polyphenol from a plant or a processed product thereof in which theobromine and polyphenol coexist.
- cocoa polyphenols contained in cocoa are known to have various physiological effects such as an antioxidant action, a calculus formation-inhibiting action, an antitumor action, an anti-stress action, and a carcinogenesis-preventing action (for example, Patent Document 1). To 5).
- Cocoa polyphenol can be obtained as an extract of cacao, and in recent years, cacao food and drink such as cocoa and chocolate using such an extract has attracted attention.
- cacao food and drink such as cocoa and chocolate using such an extract has attracted attention.
- the content of cocoa polyphenol contained in commercially available cocoa or chocolate is low. Therefore, in order to exhibit the above physiological effect by cacao food and drink, it is necessary to ingest them in a considerably large amount.
- such large intake can cause various adverse effects. Therefore, if a method for producing a composition having a high polyphenol content, particularly a composition containing a high content of cacao polyphenol, can be established, a sufficient amount of polyphenol is exerted to exert a physiological effect without ingesting a large amount of cocoa or chocolate. It can be ingested.
- JP-A-7-213251 Japanese Patent Laid-Open No. 7-238028 JP 7-274894
- a Japanese Patent Laid-Open No. 9-206026 JP-A-9-234018 Japanese Patent Laid-Open No. 9-224606 JP 2000-256345 A Martin L. Price and Larry G. Butler, J. Agric Food Chem., Vol. 25 No.6,61268-1273, 1977 AOAC Official Methods of Analysis (1990) 980.14, "Theobromine and Caffeine in Cacao Products Liquid Chromatographic Method" Food Chemistry 68 (2000) Yinrong Lu, L.
- cocoa polyphenol fraction often contains theobromine. While theobromine has an excellent physiological effect, it is known that when they are ingested in a large amount, they feel unwell. Therefore, when the cocoa polyphenol extract is used as a food or drink, it is desirable that the cocoa polyphenol content in the extract is high while the theobromine content is low.
- an object of the present invention is to provide a composition having a high polyphenol content in a simple and efficient manner, and particularly to provide a method for producing a composition having a high cocoa polyphenol content and a low theobromine content.
- the present inventors have found that theobromine is efficiently obtained when a crude extract containing polyphenol and theobromine coexists with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment. It was found that the polyphenol was adsorbed by the resin and eluted efficiently.
- the present invention is based on this finding, and has the following features.
- the manufacturing method of the polyphenol containing composition which has the following processes.
- B) The crude polyphenol extract is brought into contact with a cation exchange resin that has been subjected to a hydrogen ion replacement treatment in advance, and subsequently, a first solvent that does not contain an ionic substance is passed through the cation exchange resin.
- C The first polyphenol eluate is again passed through a cation exchange resin that has been previously subjected to hydrogen ion replacement treatment, and then a second solvent that does not contain an ionic substance is passed through to provide a second polyphenol eluate.
- a step of concentrating or drying the second polyphenol eluate which has the following processes.
- step (c) Prior to the step (c), a solution obtained by precipitating the theobromine precipitate from the first polyphenol eluate and then removing the precipitate is used in the step (c).
- the second solvent not containing the ionic substance is deionized water of 35 ° C. or lower or an aqueous ethanol solution having an ethanol concentration of 30% by weight or lower, according to any one of (1) to (6) above. Manufacturing method.
- a polyphenol-containing composition having a polyphenol content in a solid content of 33% by weight or more and a theobromine content of 1% by weight or less.
- An apparatus for producing a polyphenol-containing composition from a plant body containing polyphenol or a processed product thereof An extraction section having means for obtaining a polyphenol crude extract from a plant containing polyphenol or a processed product thereof;
- the polyphenol crude extract transferred from the extraction section via the transfer means is brought into contact with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then the first solvent that does not contain an ionic substance in the cation exchange resin.
- the first polyphenol eluate having a means for obtaining a first polyphenol eluate through which the liquid is passed, and the first polyphenol eluate transferred from the first separation / purification section via the transfer means, in advance.
- the second separation / purification having means for obtaining a second polyphenol eluate by passing a second solvent not containing an ionic substance is enabled.
- Separation and purification division A recovery section having means for concentrating or drying the second polyphenol eluate transferred from the separation and purification section via a transfer means;
- a manufacturing apparatus comprising:
- a high polyphenol-containing composition can be produced by a simple method and efficiently.
- the process since it is not necessary to pretreat the crude extract with a porous resin in advance, there are advantages that the process is simple, the polyphenol recovery rate is high, and the amount of solvent required is small.
- the composition obtained by the manufacturing method of this invention has a high polyphenol content in solid content compared with a general conventional cocoa extract, it is useful in various applications.
- the cocoa polyphenol-rich composition obtained according to the present invention has a lower theobromine content than conventional cocoa extracts, and is therefore easy to blend into foods and drinks and pharmaceuticals.
- FIG. 1 is a flowchart for explaining an embodiment of the production apparatus of the present invention.
- FIG. 2 is a graph showing the relationship between the ethanol concentration and the extraction effect when an aqueous ethanol solution is used as the extraction solvent.
- FIG. 3 is a graph showing the measurement results of micelle insolubilization ability examined in Example 6 and Comparative Example 1.
- the first of the present invention relates to a method for producing a polyphenol-containing composition.
- This production method comprises a step of extracting a polyphenol-containing plant or a processed product thereof with a solvent to obtain a polyphenol crude extract, and bringing the polyphenol crude extract into contact with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment. And then elution to obtain a polyphenol eluate and, if necessary, a step of concentrating or drying the polyphenol eluate.
- unnecessary components in the polyphenol crude extract can be removed by adsorbing to the cation exchange resin.
- a fraction rich in polyphenols can be selectively eluted.
- a preferred embodiment of the production method of the present invention includes (a) extracting a plant body containing polyphenol or a processed product thereof with a solvent to obtain a polyphenol crude extract, and (b) the polyphenol crude extract, Contacting with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and subsequently passing a first solvent that does not contain an ionic substance through the cation exchange resin to obtain a first polyphenol eluate; ) After passing the first polyphenol eluate again through a cation exchange resin that has been subjected to hydrogen ion replacement treatment in advance, the second polyphenol elution is continued by passing through a second solvent that does not contain an ionic substance.
- step (d) of concentrating or drying the second polyphenol eluate as necessary a step (d) of concentrating or drying the second polyphenol eluate as necessary.
- the cation exchange resin used in the step (c) uses the cation exchange resin prepared separately even if the cation exchange resin used in the previous step (b) is regenerated and reused. May be.
- plant or a processed product thereof containing polyphenol used in the present specification only needs to contain a polyphenol in the component of the plant or processed product thereof. Means not limited. That is, the production method according to the present invention can be applied to various plants containing polyphenols and processed products thereof. Although not particularly limited, the present invention is used to selectively extract a polyphenol component from a plant body such as cacao, mate, tea tree, cola, guarana, or coffee tree, or a processed product thereof, in which polyphenol and theobromine coexist. Useful.
- polyphenol-rich composition used in the present specification intends that the content of the polyphenol component in the solid content is 33% by weight or more, more preferably 40% by weight or more.
- the content of cacao polyphenol in the solid content is 33% by weight or more, more preferably 40% by weight or more, while theobromine in the solid content. Is intended to be 1% by weight or less, more preferably 0.9% by weight or less.
- Examples of the “cocoa plant or processed product thereof” that can be used as a raw material in the present invention include plants such as cacao bark, cacao leaves, cacao beans, and cacao shell, and cacao nibs, cacao mass, defatted cacao mass, cocoa powder, and the like.
- cocoa beans The cacao mass is obtained by grinding cacao beans, and the defatted cacao mass is obtained by removing fats and oils from the cacao mass.
- the method for removing fat from cocoa mass is not particularly limited, and a known method such as pressing can be applied.
- Cocoa powder is obtained by grinding the defatted cocoa mass. And among these exemplified raw materials, cocoa mass and cocoa powder are subjected to atomization treatment such as grinding and pulverization in the processing step, so that the extraction of polyphenols can be carried out efficiently. preferable.
- step (a) relates to a step of obtaining a polyphenol crude extract from a raw material containing polyphenol.
- This step is not particularly limited except that solvent extraction is performed, and a known extraction technique can be applied.
- a method of putting a raw material in an extraction kettle and immersing it in a predetermined amount of extraction solvent for a certain time to obtain an extract a method of sending an extraction solvent to a raw material packed in a column, and obtaining a predetermined amount of extract What is necessary is just to select suitably from a well-known solvent extraction method.
- Components (residues) that do not dissolve in the extraction solvent are removed using a method such as centrifugation or filtration.
- the polyphenol crude extract thus obtained may be a solution containing a large amount of solvent as obtained by the extraction treatment or a solution obtained by distilling a part of the extraction solvent.
- a precipitate or a precipitate is generated when a part of the extraction solvent is distilled off, it is removed by a known method such as centrifugation or filtration.
- an extract by solvent extraction (hereinafter abbreviated as “cacao crude extract”) is prepared.
- the preparation method is not particularly limited except that solvent extraction is applied.
- the solvent used for solvent extraction may be an organic solvent such as water, ethanol, methanol, and acetonitrile, or an aqueous solution thereof. Although it does not specifically limit, water, ethanol, and its aqueous solution are preferable from a viewpoint which is a solvent generally used for foodstuff manufacture. These solvents are also preferable because they are harmless to the human body.
- ethanol may be used alone, but it is more preferable to use water or a mixture of water and ethanol, that is, an aqueous ethanol solution.
- the ethanol concentration in the aqueous ethanol solution is preferably 30 to 95% by weight, more preferably 40 to 70% by weight, and still more preferably 50% by weight.
- the temperature during extraction is theoretically in the range of 0 to 100 ° C., preferably in the range of 50 to 90 ° C. when water is used as the extraction solvent.
- the temperature is theoretically in the range of 0 to 80 ° C., preferably in the range of 40 to 70 ° C.
- the extraction time may be appropriately determined according to the desired extraction object in consideration of the raw material to be used and other extraction parameters. Although not particularly limited, the extraction time is usually about 10 to 60 minutes.
- the cocoa crude extract obtained as described above contains a large amount of other unnecessary components such as theobromine, amino acids, saccharides, pigments, and fats and oils together with cocoa polyphenol.
- steps (b) and (c) relate to separation and purification using a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment.
- the purpose of step (b) is to obtain a first polyphenol eluate by mainly removing unnecessary components such as proteins and amino acids from the polyphenol crude extract obtained in step (a).
- the step (c) is intended to obtain a second polyphenol eluate by removing mainly unnecessary components such as theobromine from the eluate of the step (b). Unnecessary components are removed by bringing the cation exchange resin into contact with the crude extract or the first polyphenol eluate, and then passing an appropriate solvent therethrough.
- cation exchange resin used in the present specification is not particularly limited, and a well-known cation exchange resin having strong or weak acid is intended. Furthermore, “a cation exchange resin that has been subjected to a hydrogen ion replacement treatment in advance” indicates that the cation in the resin that is a reactive group has been previously replaced with a hydrogen ion.
- Specific examples of cation exchange resins that can be used in the present invention include Diaion (registered trademark) series SK1B, SK110, SK116, P206, WK40 manufactured by Mitsubishi Chemical Corporation, and Amberlite (registered trademark) manufactured by Rohm and Haas. IR-120B, IR-200CT, IRC50, IR-124, and Dowex (registered trademark) 50W-X8 manufactured by The Dow Chemical Company.
- the hydrogen ion replacement treatment for the cation exchange resin can be carried out, for example, by bringing an appropriate concentration of acid into contact with the resin.
- 1N-hydrochloric acid can be used to perform the above substitution treatment.
- the cation exchange resin used in the present invention is not particularly limited, but it is indispensable to perform a hydrogen ion replacement treatment in advance even when any resin is used.
- elution of a target substance from an ion exchange chromatography using a cation exchange resin or the like as a packing material includes hydrochloric acid, sodium hydroxide, calcium chloride, sodium chloride, and the like containing substances that can be ion exchanged with the ion exchange resin.
- An ionic solution is used.
- the elution of polyphenol from the cation exchange resin is carried out by passing a solvent containing no ionic substance such as deionized water.
- solvent that does not contain an ionic substance intends a solvent that does not contain a substance that adsorbs and desorbs with a cation exchange resin.
- the solvent not containing an ionic substance examples include deionized water and an aqueous solution of an organic solvent.
- the aqueous solution of the organic solvent is as exemplified above in the step (a), and a preferable solvent is an aqueous ethanol solution.
- deionized water is most preferable from the viewpoint of cost.
- an unnecessary component such as theobromine in the cacao crude extract is selectively converted into a cation exchange resin by a combination of a cation exchange resin that has been previously replaced with hydrogen ions and a solvent that does not contain an ionic substance. It is possible to adsorb and preferentially elute cocoa polyphenols. In one embodiment of the present invention, it is preferable to select and use an appropriate elution solvent according to the purposes of steps (b) and (c). Hereinafter, steps (b) and (c) will be described in more detail.
- the contact between the cation exchange resin and the polyphenol crude extract is obtained by resinizing unnecessary components other than cacao polyphenol in the crude extract into the cation exchange resin. Any method may be used as long as it can be adsorbed on the sucrose and subsequently the cocoa polyphenol fraction can be selectively eluted.
- the conditions for the contact treatment between the cacao crude extract and the cation exchange resin can be appropriately selected according to the concentration of the cacao crude extract and the like.
- step (b) of the present invention is carried out by filling a column with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then passing the cacao crude extract through the cation exchange resin in the column. Can do.
- step (b) of the present invention can also be carried out by directly adding a cation exchange resin that has been previously subjected to hydrogen ion replacement treatment to the cacao crude extract and stirring. When such a method is used, it is necessary to adsorb unnecessary components in the cacao crude extract to the cation exchange resin, and then recover the cation exchange resin by a separation method such as filtration. That is, since the latter method is batch processing, a method using a column is more preferable from the viewpoint of work efficiency.
- a solvent for equilibrating the cation exchange resin is substantially required.
- an ethanol aqueous solution having an ethanol content of 0 to 50% by weight, more preferably 0 to 10% by weight, is used as a solvent for equilibration.
- the crude cacao extract is passed through, and then the solvent containing no ionic substance is passed through to obtain the first cacao polyphenol eluate.
- the solvent used for elution in the step (b) is not particularly limited as long as it does not contain an ionic substance.
- a solvent that facilitates the separation thereof examples thereof include aqueous solutions of organic solvents such as deionized water and aqueous ethanol solution.
- the solvent used for elution is the same ethanol concentration as the extract. Therefore, for example, an ethanol aqueous solution having an ethanol content of 30 to 95% by weight can be used.
- an ethanol aqueous solution having an ethanol content of 30 to 95% by weight can be used.
- unnecessary components can be similarly adsorbed and separated on the resin.
- the temperature of the solvent used for elution is not particularly limited. For example, when deionized water is used, it is theoretically in the range of 0 to 100 ° C., preferably in the range of 5 to 50 ° C. When an aqueous ethanol solution is used, the temperature is theoretically in the range of 0 to 80 ° C., preferably in the range of 5 to 50 ° C.
- the resin amount of the cation exchange resin packed in the column can be selected as appropriate.
- the resin filling amount is preferably 1/50 times or more, more preferably 1/25 times or more the volume of the cacao crude extract.
- the resin filling amount may be reduced.
- step (c) is a step of mainly removing unnecessary components such as theobromine from the first polyphenol eluate obtained in the previous step (b).
- the cation exchange resin used in the step (c) uses the cation exchange resin prepared separately even if the cation exchange resin used in the previous step (b) is regenerated and reused. Also good.
- Theobromine is more easily adsorbed to the cation exchange resin than cocoa polyphenol. Therefore, theobromine can be removed by bringing the first cation exchange resin into contact with the first polyphenol eluate, and a polyphenol-rich fraction can be obtained.
- the method for contacting with the cation exchange resin is not particularly limited, but can be efficiently carried out by a method using a column as in the step (b). Specifically, after the cation exchange resin is equilibrated, the first polyphenol eluate is passed, and subsequently, the solvent containing no ionic substance is passed.
- the first polyphenol eluate obtained by step (b) may be used as it is.
- a solution obtained by precipitating unnecessary components from the eluate and removing the precipitate may be used.
- Such an embodiment is particularly preferable when the amount of the cation exchange resin used in step (b) is small. This is because when the amount of the cation exchange resin used in step (b) is small, the eluate contains a large amount of theobromine. Therefore, prior to step (c), theobromine contained in the first polyphenol eluate can be removed as a precipitate. Therefore, such an embodiment is beneficial for efficiently obtaining a polyphenol-rich composition having a low theobromine content.
- Theobromine precipitation can be carried out by a known method such as evaporation of the solvent in the eluate.
- the separated theobromine can be appropriately purified by a known method and used as a by-product.
- step (c) the solution obtained by removing the theobromine precipitate is brought into contact with the cation exchange resin as it is, and a solvent containing no ionic substance is passed through.
- ethanol may be appropriately added to the solution so that the ethanol concentration in the solution is approximately the same as that in the eluate.
- the solution to which ethanol has been added is brought into contact with the cation exchange resin, and a solvent containing no ionic substance is passed through.
- the solvent used for elution in the step (c) may be any solvent that can efficiently adsorb theobromine to the cation resin and can selectively elute polyphenols and does not contain an ionic substance.
- examples thereof include aqueous solutions of organic solvents such as deionized water and aqueous ethanol solution.
- the deionized water temperature is desirably 35 ° C. or lower, more preferably 25 ° C. or lower.
- theobromine is difficult to adsorb on the resin and tends to be eluted with polyphenol.
- an aqueous solution of an organic solvent such as an ethanol aqueous solution as the elution solvent
- an ethanol aqueous solution having a lower concentration than the ethanol aqueous solution used during extraction can be used.
- an aqueous ethanol solution when used in step (c), it is preferable to use an aqueous ethanol solution having an ethanol content of 0 to 30% by weight, more preferably 0 to 10% by weight.
- the ethanol concentration of the aqueous ethanol solution exceeds 30% by weight, theobromine is difficult to adsorb and tends to be eluted together with the polyphenol.
- the flow rate of the elution solvent in step (c) may be the same as in step (b) described above.
- the filling amount of the resin in the step (c) is preferably 1 or more times, preferably 1.5 or more times the amount of the first polyphenol eluate or the solution obtained by further treatment.
- the cation exchange resin used in step (b) may be regenerated and reused.
- the separation and purification using the cation exchange resin is not limited to two steps (b) and (c), and the same separation and purification may be repeated as necessary. Also good.
- a known technique relating to the separation and purification may be applied.
- concentration or drying it becomes easy to obtain a target polyphenol-containing composition in a solid state.
- concentration or drying a known method can be applied.
- concentration method include vacuum concentration and heat concentration.
- drying method include spray drying and freeze drying.
- the theobromine precipitate is generated during the concentration or drying in the step (d)
- the theobromine content in the finally obtained composition can be further reduced by removing the precipitate. .
- the removal of theobromine precipitate can be carried out by a known method such as filtration or centrifugation.
- the 2nd of this invention is related with the polyphenol high content composition obtained by the manufacturing method by the 1st of this invention. Since such a composition has a high polyphenol content per solid content, it can be used for various applications in which the polyphenol is effective.
- the cocoa polyphenol composition obtained by the present invention is characterized in that the cocoa polyphenol content is high, while the theobromine content is significantly lower than that of conventional products. Therefore, it can add suitably to food-drinks and a pharmaceutical.
- compositions such as tablets, capsules, granules, powders and the like having a pharmacological effect are provided by blending the composition according to the present invention with various excipients, stabilizers, flavors and the like that can be generally used in the preparation. be able to.
- the composition according to the present invention can be used for food and drink such as cocoa, coffee, chocolate, biscuits, snacks, candy, tablet confectionery, gum, gummy, jelly, sheep cane, ice cream, sorbet, beverage, dairy product, bread, sausage and ham.
- Pharmacological effects can be imparted to them by blending them into the product.
- a cocoa polyphenol-containing composition having a polyphenol content in a solid content of 33% by weight or more and a theobromine content of 1% by weight or less is preferable. According to the present invention, a desirable composition having a high cocoa polyphenol content and a low theobromine content can be provided.
- the 3rd of this invention is related with the apparatus for manufacturing a polyphenol containing composition from the plant body containing polyphenol, or its processed goods.
- the production apparatus of the present invention includes an extraction section having means for obtaining a polyphenol crude extract from a plant body containing polyphenol or a processed product thereof, and a polyphenol crude extract transferred from the extraction section via a transfer means in advance.
- a first cation exchange resin having a means for obtaining a first polyphenol eluate by contacting the cation exchange resin that has been subjected to hydrogen ion substitution treatment, and subsequently passing a first solvent that does not contain an ionic substance through the cation exchange resin;
- the first polyphenol eluate transferred from the first separation / purification section through the transfer means is again passed through a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then contains an ionic substance.
- a separation / purification section enabling a second separation / purification having means for obtaining a second polyphenol eluate by passing a second solvent not present, and the separation Recovery section and having a means for concentrating or drying the second polyphenol eluate transferred through the transfer means from the manufacturing division It is characterized by having.
- the extraction section includes at least a container that can contain the plant body or a processed product thereof (hereinafter also referred to as a raw material) and a solvent used for extraction.
- the extraction is carried out by bringing the raw material and the solvent into contact with each other in the container, and the insoluble component (residue) is removed by the removing means to obtain a polyphenol crude extract.
- the container preferably includes at least one opening for injecting the raw material and the extraction solvent and an opening for discharging the polyphenol crude extract.
- the container may be a column, an extraction kettle, or a tank.
- stirring means such as a propeller and a screw may be provided in the container.
- a means for finely pulverizing the raw material such as a rotary blade may be provided in the container.
- the insoluble component removing means may be, for example, a centrifuge device provided outside the container and a filtration device. If necessary, by adding a concentration means such as an evaporator or a vacuum concentrator to the extraction section, it is possible to distill off the solvent of the crude extract prior to transfer to the separation and purification section, thereby reducing the liquid volume.
- the polyphenol crude extract obtained in the extraction section is transferred to the separation and purification section through a transfer means.
- the transfer means may be a liquid feed pump and a pipe, for example. If necessary, a control valve may be provided in the pipe. The flow rate and flow rate when the polyphenol crude extract is transferred to the separation and purification section can be adjusted by the liquid feed pump and the control valve.
- the separation and purification section has an arrangement that enables at least two separation and purification steps including the first and second separation and purification. That is, the first and second separation and purification may be arranged as independent sections, or may be arranged with transfer means such as piping that enables circulation within the same section.
- the first separation and purification section includes a first column containing a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and an ionic substance.
- a first solvent tank that contains a first solvent not included is provided. After passing the polyphenol crude extract transferred from the extraction section through the resin contained in the column, the first solvent not containing an ionic substance transferred from the first solvent tank is further passed. As a result, a first polyphenol eluate is obtained.
- the second separation and purification section includes a second column containing a cation exchange resin that has been previously subjected to hydrogen ion replacement treatment, and a second solvent tank that contains a second solvent that does not contain an ionic substance. .
- the second solvent not containing an ionic substance transferred from the second solvent tank is further passed.
- a second polyphenol eluate is obtained.
- the first separation / purification section and the second separation / purification section are communicated with each other through a transfer means such as a pipe, and the first polyphenol eluate obtained in the first separation / purification section is connected with the second through the transfer means. To the separation and purification section.
- a removal section for removing theobromine may be provided before the first polyphenol eluate is transferred to the second separation and purification section.
- a removal section is, for example, provided between the first and second separation and purification sections, a container for storing the first polyphenol eluate, means for precipitating and removing theobromine from the eluate, and theobromine.
- a transfer means such as a pipe for transferring the solution obtained after the removal to the second separation and purification section is provided.
- the removing means may be, for example, a filtration device or a centrifuge device.
- the removal section may be provided with a temperature control means such as a cooling device and a concentrating device such as an evaporator or a vacuum concentrator in order to promote precipitation of theobromine.
- the separation and purification section includes a column containing a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and a first that does not contain an ionic substance.
- a transfer means such as piping for transferring to the column is provided.
- the polyphenol crude extract transferred from the extraction section is passed through the column, and further, the first solvent supplied from the first solvent tank is passed through, so that the first A polyphenol eluate is obtained.
- the first polyphenol eluate is transferred again to the regenerated column via the transfer means, and after passing through the column, the second solvent supplied from the second solvent tank is passed. As a result, a second polyphenol eluate is obtained.
- the separation / purification section may include third and fourth solvent tanks that contain treatment solutions such as hydrochloric acid and sodium hydroxide solution used for washing the resin to perform cation exchange resin hydrogen substitution treatment. Furthermore, you may add the solvent tank which accommodates the solvent supplied to a column as needed. After obtaining the polyphenol eluate, the treatment solution is passed through the column from the third and fourth solvent tanks, whereby the resin in the column is regenerated and separation and purification can be carried out continuously.
- the supply of the solvent from the first to third solvent tanks to the column is achieved by transfer means such as a liquid feed pump and piping. This transfer means may include a regulating valve as required. The flow rate and flow rate of the solvent supplied to the column can be adjusted by the liquid feed pump and the control valve.
- the polyphenol eluate obtained in the separation / purification section is transferred to the collection section through a transfer means.
- the transfer means may be a liquid feed pump and a pipe, for example. If necessary, a control valve may be provided in the pipe. The flow rate and flow rate when the polyphenol crude extract is transferred to the collection section can be adjusted by the feed pump and the control valve.
- the polyphenol eluate is transferred to the collection section through the transfer means.
- the collection section has a container for storing the transferred polyphenol eluate and means for removing the solvent in the eluate from the container.
- the means for removing the solvent may be an apparatus used for ordinary pulverization such as ordinary concentration such as a vacuum concentrator, freeze dryer, or spray dryer.
- FIG. 1 shows an embodiment of the manufacturing apparatus of the present invention.
- the manufacturing apparatus of the present invention has an extraction section 100, a separation and purification section 200, and a recovery section 300.
- the extraction section 100 includes an extraction tank 110 for carrying out extraction from raw materials, a raw material tank 120 for containing raw materials, a solvent tank 130 for containing extraction solvents, and a centrifuge for removing insoluble matters after extraction. 140.
- the extraction tank 110 and the centrifuge 140 are communicated with each other by a pipe 100a.
- the separation / purification section 200 contains a column 210 for performing the first separation / purification, a concentrator 220 for concentrating the eluate, a column 230 for performing the second separation / purification, and a solvent for performing the column washing process.
- a solvent tank 240a for storing the solvent used for the column equilibration process, and solvent tanks 240c and 240d for storing the solvent used for elution.
- the column 210, the concentrator 220, and the column 230 communicate with each other via pipes 200a and 200b, respectively.
- Such a separation / purification section 200 communicates with the extraction section 100 via a pipe 100b.
- Reference numerals 212 and 232 denote waste liquid tanks for storing waste liquid, respectively.
- the recovery section 300 has a concentrator 310 for concentrating the eluate transferred from the separation and purification section 200 via the pipe 200c, and a spray dryer 320 for drying the concentrated liquid.
- the concentrator 310 and the spray dryer 320 communicate with each other via a pipe 300a.
- Reference numeral 322 denotes a recovery tank containing a composition obtained by drying.
- a composition having a high polyphenol content can be efficiently obtained using a plant body containing polyphenol or a processed product thereof as a raw material.
- the manufacturing apparatus of the present invention is intended to be used for carrying out the manufacturing method of the present invention described above. Therefore, various conditions such as the solvent used in each section and the resin accommodated in the column are the same as those in the production method.
- cocoa polyphenol content and theobromine content described in each Example and each Comparative Example are values obtained by measurement by the following methods.
- the polyphenol content was measured by the Prussian blue method. More specifically, referring to the method described in Martin L. Price and Larry G. Butler, J. Agric Food Chem., Vol. 25 No. 6, 1268-1273, 1977, commercially available epicatechin was used as a standard product. The values obtained by quantitative analysis of the solid content of the compositions obtained in the examples and comparative examples.
- the outline of a specific analysis method is as follows.
- the composition was degreased by treatment with n-hexane, and then extracted with a 50 wt% aqueous methanol solution to obtain a test solution.
- 100 ⁇ L of each test solution was added to 50 mL of distilled water, and 3 mL of 0.1 M iron (III) ammonium sulfate-0.1N hydrochloric acid solution was added with stirring. After 20 minutes, 3 mL of 8 mM hexacyanoiron (III) potassium aqueous solution was added.
- the absorbance at 720 nm was measured after another 20 minutes.
- Theobromine content was obtained in each Example and each Comparative Example with reference to the method described in AOAC Official Methods of Analysis (1990) 980.14 ⁇ Theobromine and Caffeine in Cacao Products Liquid Chromatographic Method '' and using commercially available theobromine as a standard product. It is the value obtained by performing a quantitative analysis about solid content of the composition obtained.
- the outline of a specific analysis method is as follows. First, accurately weigh the composition as a sample in a centrifuge tube, add 30 ml of petroleum ether and stir well, then centrifuge and discard the supernatant. Transfer the degreased sample into an Erlenmeyer flask and add water to make about 100 ml. This solution was heated in a 100 ° C. water bath for 25 minutes. The heated solution was immediately cooled, 10 ml of a 2 wt% aqueous zinc sulfate solution and 10 ml of 1.8 wt% barium hydroxide were added and mixed, and then allowed to stand. Water was added to this solution to make up to 200 ml, and the solution was heated again in a 100 ° C.
- a cation exchange resin Amberlite IR-120B
- a cation exchange resin Dowex 50W-X8
- a cation exchange resin Amberlite IR-120B
- a cation exchange resin Amberlite IR-120B
- deionized water was passed through the column.
- 1500 ml of the eluate was collected.
- 1500 ml of the eluate was freeze-dried to obtain a polyphenol-containing composition.
- the polyphenol content was 20.5% by weight and the theobromine content was 0.4% by weight.
- the cacao crude extract separately prepared was freeze-dried and analyzed for components in the solid content. As a result, the polyphenol content was 7.8% by weight and the theobromine content was 0.9% by weight.
- a cation exchange resin Amberlite IR-120B
- Example 6 Using the polyphenol-containing composition obtained in Example 5, the physiological effects of the composition were examined from the following evaluation items. The results are shown in Table 1 and FIG.
- DPPH radical scavenging ability DPPH (1,1-Dyphenyl-2-picrylhydrazyl; DPPH) radical scavenging ability is reported in Food Chemistry 68 (2000) Yinrong Lu, L. Yeap Foo “Antioxidant and radical scavenging According to the method described in "Activities of polyphenols from apple pomace", measurement was performed using commercially available epicatechin as an index. The index epicatechin and each sample were dissolved in 50 wt% aqueous ethanol solution. To 100 ⁇ l of this solution, 2 ml of 0.1 mM DPPH solution was added and allowed to stand at room temperature for 30 minutes, and then the absorbance at 520 nm was measured. The inhibition rate of each sample was calculated using the absorbance value of the blank solution (DPPH solution) as the inhibition rate of 100%. It shows that DPPH radical scavenging ability is so high that the numerical value obtained by the measurement is low.
- Example 1 A commercially available cocoa polyphenol material (manufactured by Meiji Food Materia Co., Ltd., trade name “cocoa polyphenol”) was used, and the same examination as in Example 6 was performed. The results are shown in Table 1 and FIG.
- Each content is a content rate based on the total weight of the solid content.
- the polyphenol-containing composition of the present invention (Example 6) has a higher polyphenol content and a lower theobromine content than the commercially available product (Comparative Example 1). Moreover, it turns out that radical scavenging ability is high compared with a commercial item. Furthermore, it can be seen from FIG. 3 that the micelle insolubility is higher than that of the commercially available product.
- “high DPPH radical scavenging ability” suggests that the composition has high antioxidant power.
- “high micelle insolubilizing ability” suggests that cholesterol has a high ability to insolubilize micelles and has an ability to lower blood cholesterol. From such a viewpoint, it is apparent that the present invention can provide a more useful polyphenol-containing composition than conventional commercial products.
- Example 7 This example relates to examination of a cation exchange resin used in the separation and purification process.
- a cocoa bean (dried in the production area, cocoa polyphenol content 7.0% by weight, theobromine content 1.2% by weight) was used, and a polyphenol-containing composition was prepared according to the following procedure. First, after crushing 10 g of cocoa beans, 200 ml of 50 wt% ethanol aqueous solution was added and stirred at 50 ° C. for 30 minutes, and then insoluble components were removed by centrifugation to obtain a cacao crude extract. .
- Example 2 By performing the same operation as in Example 7, except that instead of the various cation exchange resins used in Example 7, an anion exchange resin (see Table 2) subjected to hydroxide ion substitution treatment was used. The eluate was obtained. Next, the eluate was freeze-dried in the same manner as in Example to obtain a polyphenol-containing composition. The components in the solid content of the obtained composition were analyzed in the same manner as in Example 7. The results are shown in Table 2.
- Each content is a content rate based on the total weight of the solid content.
- the “Diaion” (registered trademark) series is manufactured by Mitsubishi Chemical Corporation.
- a series of “Amberlite” (registered trademark) is manufactured by Rohm and Haas Co., Ltd.
- Example 8 This example relates to the examination of the solvent used in the process of separation and purification using a cation exchange resin. Specifically, various types of solvents were used as the equilibration solution for the column packed with the cation exchange resin and the eluent from the column. A cocoa bean (dried in the production area, cocoa polyphenol content 7.4% by weight, theobromine content 1.3% by weight) was used, and a polyphenol-containing composition was prepared according to the following procedure. First, after grinding 2 g of cocoa beans, 20 ml of 50 wt% aqueous ethanol solution was added, stirred at 50 ° C. for 30 minutes, and then insoluble components were removed by centrifugation to obtain a crude cocoa extract. .
- the temperature of the equilibration solution is 25 ° C.
- the temperature of the column eluent is 25 ° C.
- Each content is a content rate based on the total weight of the solid content.
- a cation exchange resin Amberlite IR-120B strong acid cation exchange resin
- a cation exchange resin Amberlite IR-120B strong acid cation exchange resin
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Abstract
Description
(a)ポリフェノールを含有する植物体またはその加工品を溶媒で抽出し、ポリフェノール粗抽出液を得る工程、
(b)上記ポリフェノール粗抽出液を、予め水素イオン置換処理した陽イオン交換樹脂に接触させ、引き続き上記陽イオン交換樹脂に、イオン性物質を含まない第1の溶媒を通液し、第1のポリフェノール溶出液を得る工程、
(c)上記第1のポリフェノール溶出液を、予め水素イオン置換処理した陽イオン交換樹脂に再度通液した後に、イオン性物質を含まない第2の溶媒を通液し、第2のポリフェノール溶出液を得る工程、および
(d)上記第2のポリフェノール溶出液を濃縮または乾燥させる工程。 (1) The manufacturing method of the polyphenol containing composition which has the following processes.
(A) extracting a plant body containing polyphenol or a processed product thereof with a solvent to obtain a polyphenol crude extract,
(B) The crude polyphenol extract is brought into contact with a cation exchange resin that has been subjected to a hydrogen ion replacement treatment in advance, and subsequently, a first solvent that does not contain an ionic substance is passed through the cation exchange resin. Obtaining a polyphenol eluate,
(C) The first polyphenol eluate is again passed through a cation exchange resin that has been previously subjected to hydrogen ion replacement treatment, and then a second solvent that does not contain an ionic substance is passed through to provide a second polyphenol eluate. And (d) a step of concentrating or drying the second polyphenol eluate.
ポリフェノールを含有する植物体またはその加工品からポリフェノール粗抽出液を得る手段を有する抽出区分と、
上記抽出区分から移送手段を介して移送されたポリフェノール粗抽出液を、予め水素イオン置換処理した陽イオン交換樹脂に接触させ、引き続き上記陽イオン交換樹脂に、イオン性物質を含まない第1の溶媒を通液し、第1のポリフェノール溶出液を得る手段を有する第1の分離精製、および上記第1の分離精製区分から移送手段を介して移送された上記第1のポリフェノール溶出液を、予め水素イオン置換処理した陽イオン交換樹脂に再度通液した後に、イオン性物質を含まない第2の溶媒を通液して第2のポリフェノール溶出液を得る手段を有する第2の分離精製を可能とする分離精製区分と、
上記分離精製区分から移送手段を介して移送された上記第2のポリフェノール溶出液を濃縮または乾燥する手段を有する回収区分と
を有することを特徴とする製造装置。 (10) An apparatus for producing a polyphenol-containing composition from a plant body containing polyphenol or a processed product thereof,
An extraction section having means for obtaining a polyphenol crude extract from a plant containing polyphenol or a processed product thereof;
The polyphenol crude extract transferred from the extraction section via the transfer means is brought into contact with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then the first solvent that does not contain an ionic substance in the cation exchange resin. The first polyphenol eluate having a means for obtaining a first polyphenol eluate through which the liquid is passed, and the first polyphenol eluate transferred from the first separation / purification section via the transfer means, in advance. After passing again through the ion-exchange-treated cation exchange resin, the second separation / purification having means for obtaining a second polyphenol eluate by passing a second solvent not containing an ionic substance is enabled. Separation and purification division,
A recovery section having means for concentrating or drying the second polyphenol eluate transferred from the separation and purification section via a transfer means;
A manufacturing apparatus comprising:
100a、100b:配管
110:抽出タンク
120:原料タンク
130:溶媒タンク
140:遠心分離機
200:分離精製区分
200a、200b、200c:配管
210:カラム
212:廃液タンク
220:濃縮機
230:カラム
232:廃液タンク
240a、240b、240c、240d:溶媒タンク
300 回収区分
300a 配管
310:濃縮機
320:噴霧乾燥機
322: 回収タンク 100:
本発明の製造装置は、ポリフェノールを含有する植物体またはその加工品からポリフェノール粗抽出液を得る手段を有する抽出区分と、上記抽出区分から移送手段を介して移送されたポリフェノール粗抽出液を、予め水素イオン置換処理した陽イオン交換樹脂に接触させ、引き続き上記陽イオン交換樹脂に、イオン性物質を含まない第1の溶媒を通液し、第1のポリフェノール溶出液を得る手段を有する第1の分離精製および上記第1の分離精製区分から移送手段を介して移送された上記第1のポリフェノール溶出液を、予め水素イオン置換処理した陽イオン交換樹脂に再度通液した後に、イオン性物質を含まない第2の溶媒を通液して第2のポリフェノール溶出液を得る手段を有する第2の分離精製を可能とする分離精製区分と、上記分離精製区分から移送手段を介して移送された上記第2のポリフェノール溶出液を濃縮または乾燥する手段を有する回収区分と
を有することを特徴とする。 3rd of this invention is related with the apparatus for manufacturing a polyphenol containing composition from the plant body containing polyphenol, or its processed goods.
The production apparatus of the present invention includes an extraction section having means for obtaining a polyphenol crude extract from a plant body containing polyphenol or a processed product thereof, and a polyphenol crude extract transferred from the extraction section via a transfer means in advance. A first cation exchange resin having a means for obtaining a first polyphenol eluate by contacting the cation exchange resin that has been subjected to hydrogen ion substitution treatment, and subsequently passing a first solvent that does not contain an ionic substance through the cation exchange resin; The first polyphenol eluate transferred from the first separation / purification section through the transfer means is again passed through a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then contains an ionic substance. A separation / purification section enabling a second separation / purification having means for obtaining a second polyphenol eluate by passing a second solvent not present, and the separation Recovery section and having a means for concentrating or drying the second polyphenol eluate transferred through the transfer means from the manufacturing division
It is characterized by having.
ポリフェノール含量は、プルシアンブルー法によって測定した。より詳細には、Martin L. Price and Larry G. Butler, J. Agric Food Chem., Vol. 25 No.6, 1268-1273, 1977に記載の方法を参照し、市販のエピカテキンを標準品として、各実施例および各比較例で得た組成物の固形分について定量分析を行うことによって得た値である。 (Polyphenol content)
The polyphenol content was measured by the Prussian blue method. More specifically, referring to the method described in Martin L. Price and Larry G. Butler, J. Agric Food Chem., Vol. 25 No. 6, 1268-1273, 1977, commercially available epicatechin was used as a standard product. The values obtained by quantitative analysis of the solid content of the compositions obtained in the examples and comparative examples.
組成物をn-ヘキサンで処理して脱脂した後、50重量%メタノール水溶液を用いて抽出し、試験溶液とした。次いで、蒸留水50mLに各試験溶液100μLを添加し、攪拌しながら0.1M 硫酸鉄(III)アンモニウム-0.1N 塩酸溶液3mLを加え、その20分後に8mM ヘキサシアノ鉄(III)カリウム水溶液3mLを加え、さらに20分後に720nmの吸光度を測定した。さらに、蒸留水50mLに各サンプル液の溶媒(50重量%メタノール)100μLを加えたもの、および(-)エピカテキン(シグマ社製)メタノール溶液を用い、同様の処理を行って検量線を作成し、各試験溶液中のポリフェノール含量を算出した。 The outline of a specific analysis method is as follows.
The composition was degreased by treatment with n-hexane, and then extracted with a 50 wt% aqueous methanol solution to obtain a test solution. Next, 100 μL of each test solution was added to 50 mL of distilled water, and 3 mL of 0.1 M iron (III) ammonium sulfate-0.1N hydrochloric acid solution was added with stirring. After 20 minutes, 3 mL of 8 mM hexacyanoiron (III) potassium aqueous solution was added. In addition, the absorbance at 720 nm was measured after another 20 minutes. Further, using a solution obtained by adding 100 μL of the solvent of each sample solution (50 wt% methanol) to 50 mL of distilled water and a methanol solution of (−) epicatechin (manufactured by Sigma), the same treatment was performed to prepare a calibration curve. The polyphenol content in each test solution was calculated.
テオブロミン含量は、AOAC Official Methods of Analysis (1990) 980.14 「Theobromine and Caffeine in Cacao Products Liquid Chromatographic Method」に記載された方法を参照し、市販のテオブロミンを標準品として、各実施例および各比較例で得られる組成物の固形分について定量分析を行うことによって得た値である。 (Theobromine content)
Theobromine content was obtained in each Example and each Comparative Example with reference to the method described in AOAC Official Methods of Analysis (1990) 980.14 `` Theobromine and Caffeine in Cacao Products Liquid Chromatographic Method '' and using commercially available theobromine as a standard product. It is the value obtained by performing a quantitative analysis about solid content of the composition obtained.
先ず、組成物を試料として遠沈管に正確に量り取り、石油エーテル30mlを加えてよく撹拌した後、遠心分離して上清を捨てる。脱脂された試料を三角フラスコに移し入れ、水を加えて約100mlとする。この溶液を100℃の湯浴中で25分間加熱した。加熱後の溶液を直ちに冷却し、2重量%硫酸亜鉛水溶液10ml、および1.8重量%水酸化バリウム10mlを加えて混合した後、静置した。この溶液に水を加えて200mlに定容し、再度、100℃の湯浴中で10分間加熱した。加熱後の溶液をろ過することによって、試料溶液を調製した。このようにして得た試料溶液を使用して、高速液体クロマトグラフによる測定を実施した。高速液体クロマトグラフによる測定条件は、以下のとおりである。
分析用カラム: Waters μ-Bondapak C18 10μm 4mmID×300mm(またはこれと同等のもの)
移動層: 水:アセトニトリル(85:15)
移動層流量: 1.0ml/分、検出;UV273nm The outline of a specific analysis method is as follows.
First, accurately weigh the composition as a sample in a centrifuge tube, add 30 ml of petroleum ether and stir well, then centrifuge and discard the supernatant. Transfer the degreased sample into an Erlenmeyer flask and add water to make about 100 ml. This solution was heated in a 100 ° C. water bath for 25 minutes. The heated solution was immediately cooled, 10 ml of a 2 wt% aqueous zinc sulfate solution and 10 ml of 1.8 wt% barium hydroxide were added and mixed, and then allowed to stand. Water was added to this solution to make up to 200 ml, and the solution was heated again in a 100 ° C. hot water bath for 10 minutes. A sample solution was prepared by filtering the heated solution. Using the sample solution thus obtained, measurement by a high performance liquid chromatograph was performed. The measurement conditions by the high performance liquid chromatograph are as follows.
Analytical column: Waters μ-
Moving bed: water: acetonitrile (85:15)
Moving bed flow rate: 1.0 ml / min, detection; UV 273 nm
抽出溶媒としてエタノール水溶液を使用し、溶媒の組成が、ポリフェノール回収率と固形分あたりのポリフェノール含量に与える影響について検討した。手順は以下のとおりである。
先ず、10gのカカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.1重量%、テオブロミン含量1.3重量%)を磨砕した。この磨砕したカカオ豆を、0~100重量%のエタノール水溶液200mlに添加し、50℃、30分間撹拌した。次いで、不溶物を遠心分離処理にて取り除いて、カカオ粗抽出液を得た。得られたカカオ粗抽出液を凍結乾燥し、この固形分中のポリフェノール含量を測定した。結果を図2に示す。ポリフェノール回収率、および固形分あたりのポリフェノール含量の結果を考慮すると、エタノール濃度50重量%付近が抽出溶媒として好ましいことが分かった。 (Examination of extraction solvent)
An ethanol aqueous solution was used as the extraction solvent, and the influence of the solvent composition on the polyphenol recovery rate and the polyphenol content per solid content was examined. The procedure is as follows.
First, 10 g of cocoa beans (dried in the production area, cocoa polyphenol content 7.1% by weight, theobromine content 1.3% by weight) were ground. The ground cocoa beans were added to 200 ml of a 0-100 wt% aqueous ethanol solution and stirred at 50 ° C. for 30 minutes. Subsequently, insoluble matters were removed by centrifugation, and a cacao crude extract was obtained. The obtained cacao crude extract was freeze-dried, and the polyphenol content in this solid content was measured. The results are shown in FIG. Considering the result of polyphenol recovery rate and polyphenol content per solid content, it was found that an ethanol concentration of around 50% by weight is preferable as the extraction solvent.
ココアパウダー(油脂含量12重量%、カカオポリフェノール含量10.4重量%、テオブロミン含量2.0重量%)を使用し、以下の手順に従って、ポリフェノール含有組成物を調製した。
先ず、上記ココアパウダー100gを1000mlの水に分散し、90℃で30分間撹拌した後、不溶物を遠心分離処理にて取り除くことにより、カカオ粗抽出液を得た。
次に、予め水素イオン置換処理を施した陽イオン交換樹脂(アンバーライト IR-120B)を1000ml充填したカラムに、上記カカオ粗抽出液をSV=5の流速で通液した。続いて、上記カラムに25℃の脱イオン水を通液した。上記カカオ粗抽出液および上記脱イオン水の通液によって、溶出液2000mlを分取した。
次に、上記溶出液2000mlを凍結乾燥することによって、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は40.2重量%、テオブロミン含量は0.6重量%であった。
同様にして別途調製したカカオ粗抽出液を凍結乾燥し、固形分中の成分を分析した結果、ポリフェノール含量は8.2重量%、テオブロミン含量は3.3重量%であった。 Example 1
Using cocoa powder (oil content 12% by weight, cacao polyphenol content 10.4% by weight, theobromine content 2.0% by weight), a polyphenol-containing composition was prepared according to the following procedure.
First, 100 g of the cocoa powder was dispersed in 1000 ml of water, stirred at 90 ° C. for 30 minutes, and then insoluble matters were removed by centrifugal separation to obtain a crude cacao extract.
Next, the cacao crude extract was passed at a flow rate of SV = 5 through a column packed with 1000 ml of a cation exchange resin (Amberlite IR-120B) that had been previously subjected to hydrogen ion replacement treatment. Subsequently, deionized water at 25 ° C. was passed through the column. By passing the cacao crude extract and the deionized water, 2000 ml of eluate was collected.
Next, 2000 ml of the eluate was freeze-dried to obtain a polyphenol-containing composition. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 40.2% by weight and the theobromine content was 0.6% by weight.
Similarly, a separately prepared crude cacao extract was freeze-dried and analyzed for components in the solid content. As a result, the polyphenol content was 8.2% by weight and the theobromine content was 3.3% by weight.
ココアパウダー(油脂含量12重量%、カカオポリフェノール含量10.4重量%、テオブロミン含量2.0重量%)を使用し、以下の手順に従って、ポリフェノール含有組成物を調製した。
先ず、上記ココアパウダー2gを50重量%エタノール水溶液20mlに分散し、50℃で30分間撹拌した後、不溶物を遠心分離処理にて取り除くことにより、カカオ粗抽出液を得た。
次に、予め水素イオン置換処理を施した陽イオン交換樹脂(ダウエックス 50W-X8)を20ml充填したカラムに、このカカオ粗抽出液をSV=5の流速で通液した。続いて、カラムに25℃の脱イオン水を通液した。上記カカオ粗抽出液および上記脱イオン水の通液によって、溶出液50mlを分取した。
次に、上記溶出液50mlを凍結乾燥することによって、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は53.4重量%、テオブロミン含量は0.7重量%であった。
同様にして別途調製したカカオ粗抽出液を凍結乾燥し、固形分中の成分を分析した結果、ポリフェノール含量は19.9重量%、テオブロミン含量は5.3重量%であった。 (Example 2)
Using cocoa powder (oil content 12% by weight, cacao polyphenol content 10.4% by weight, theobromine content 2.0% by weight), a polyphenol-containing composition was prepared according to the following procedure.
First, 2 g of the above cocoa powder was dispersed in 20 ml of a 50 wt% ethanol aqueous solution, stirred at 50 ° C. for 30 minutes, and then insoluble matters were removed by centrifugal separation to obtain a crude cacao extract.
Next, this cacao crude extract was passed at a flow rate of SV = 5 through a column packed with 20 ml of a cation exchange resin (Dowex 50W-X8) that had been previously subjected to hydrogen ion replacement treatment. Subsequently, 25 ° C. deionized water was passed through the column. 50 ml of the eluate was collected by passing the cacao crude extract and the deionized water.
Next, 50 ml of the eluate was freeze-dried to obtain a polyphenol-containing composition. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 53.4% by weight and the theobromine content was 0.7% by weight.
Similarly, a separately prepared cacao crude extract was freeze-dried and analyzed for components in the solid content. As a result, the polyphenol content was 19.9% by weight and the theobromine content was 5.3% by weight.
10gのカカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.3重量%、テオブロミン含量1.3重量%)を使用し、以下の手順に従って、ポリフェノール含有組成物を調製した。
先ず、上記カカオ豆10gを磨砕した後、50重量%エタノール水溶液200mlを添加し、50℃にて30分間撹拌した。次いで、不溶成分を遠心分離処理にて除去することにより、カカオ粗抽出液を得た。
次に、予め水素イオン置換処理を施した陽イオン交換樹脂(アンバーライト IR-120B)を200ml充填したカラムに、上記カカオ粗抽出液をSV=5の流速で通液した。続いて、カラムに25℃の脱イオン水を通液した。上記カカオ粗抽出液および上記脱イオン水を通液によって、溶出液600mlを分取した。
次に、上記600mlの溶出液を凍結乾燥することによって、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は62.3重量%、テオブロミン含量は0.6重量%であった。
同様にして別途調製したカカオ粗抽出液を凍結乾燥し、固形分中の成分を分析した結果、ポリフェノール含量は28.4重量%、テオブロミン含量は8.5重量%であった。 (Example 3)
A polyphenol-containing composition was prepared according to the following procedure using 10 g of cocoa beans (dried in the production area, cocoa polyphenol content 7.3% by weight, theobromine content 1.3% by weight).
First, after crushing 10 g of the above cocoa beans, 200 ml of 50 wt% aqueous ethanol solution was added and stirred at 50 ° C. for 30 minutes. Subsequently, the cacao crude extract was obtained by removing insoluble components by centrifugation.
Next, the cacao crude extract was passed through a column packed with 200 ml of a cation exchange resin (Amberlite IR-120B) that had been previously subjected to hydrogen ion replacement at a flow rate of SV = 5. Subsequently, 25 ° C. deionized water was passed through the column. 600 ml of the eluate was fractionated by passing the cacao crude extract and the deionized water through.
Next, the polyphenol containing composition was obtained by freeze-drying the 600 ml eluate. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 62.3% by weight and the theobromine content was 0.6% by weight.
Similarly, a crude cacao extract separately prepared was freeze-dried and analyzed for components in the solid content. As a result, the polyphenol content was 28.4% by weight and the theobromine content was 8.5% by weight.
100gのカカオシェル(カカオポリフェノール含量3.5重量%、テオブロミン含量2.0重量%)を使用し、以下の手順に従って、ポリフェノール含有組成物を調製した。
先ず、上記カカオシェル100gを磨砕した後、50重量%エタノール水溶液500mlを添加し、50℃にて30分間撹拌した。次いで、不溶成分を遠心分離処理にて除去することにより、カカオシェル粗抽出液を得た。
次に、予め水素イオン置換処理を施した陽イオン交換樹脂(アンバーライト IR-120B)を500ml充填したカラムに、このカカオシェル粗抽出液をSV=5の流速で通液した。続いて、カラムに25℃の脱イオン水を通液した。カカオシェル粗抽出液および脱イオン水の通液によって、溶出液1500mlを分取した。
次に、上記溶出液1500mlを凍結乾燥することによって、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は20.5重量%、テオブロミン含量は0.4重量%であった。
同様にして別途調製したカカオ粗抽出液を凍結乾燥し、固形分中の成分を分析した結果、ポリフェノール含量は7.8重量%、テオブロミン含量は0.9重量%であった。 Example 4
Using 100 g of cocoa shell (cocoa polyphenol content 3.5 wt%, theobromine content 2.0 wt%), a polyphenol-containing composition was prepared according to the following procedure.
First, after crushing 100 g of the cocoa shell, 500 ml of 50 wt% aqueous ethanol solution was added and stirred at 50 ° C. for 30 minutes. Next, a cacao shell crude extract was obtained by removing insoluble components by centrifugation.
Next, this cacao shell crude extract was passed at a flow rate of SV = 5 through a column packed with 500 ml of a cation exchange resin (Amberlite IR-120B) that had been previously subjected to hydrogen ion replacement treatment. Subsequently, 25 ° C. deionized water was passed through the column. By passing the cacao shell crude extract and deionized water, 1500 ml of the eluate was collected.
Next, 1500 ml of the eluate was freeze-dried to obtain a polyphenol-containing composition. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 20.5% by weight and the theobromine content was 0.4% by weight.
Similarly, the cacao crude extract separately prepared was freeze-dried and analyzed for components in the solid content. As a result, the polyphenol content was 7.8% by weight and the theobromine content was 0.9% by weight.
100gのカカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.1重量%、テオブロミン含量1.3重量%)を使用し、以下の手順に従って、ポリフェノール含有組成物を調製した。
先ず、上記カカオ豆100gを磨砕した後、50重量%エタノール水溶液1000mlを添加し、50℃にて10分間撹拌した。次いで、不溶成分を遠心分離処理にて除去し、カカオ粗抽出液を得た。
次に、予め水素イオン置換処理を施した陽イオン交換樹脂(アンバーライト IR-120B)を1000ml充填したカラムに、上記カカオ粗抽出液をSV=5の流速で通液した。続いて25℃の脱イオン水を通液した。上記カカオ粗抽出液および上記脱イオン水の通液によって、溶出液3000mlを分取した。
次に、上記3000mlの溶出液を、乾燥温度(入口温度)160℃の条件下で噴霧乾燥を行い、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は59.0重量%、テオブロミン含量は0.6重量%であった。
同様にして別途調製したカカオ粗抽出液を凍結乾燥し、固形分中の成分を分析した結果、ポリフェノール含量は28.4重量%、テオブロミン含量は8.5重量%であった。 (Example 5)
A polyphenol-containing composition was prepared according to the following procedure using 100 g of cocoa beans (dried in the production area, cocoa polyphenol content 7.1% by weight, theobromine content 1.3% by weight).
First, after crushing 100 g of the above cocoa beans, 1000 ml of 50 wt% aqueous ethanol solution was added and stirred at 50 ° C. for 10 minutes. Next, insoluble components were removed by centrifugation, and a cacao crude extract was obtained.
Next, the cacao crude extract was passed at a flow rate of SV = 5 through a column packed with 1000 ml of a cation exchange resin (Amberlite IR-120B) that had been previously subjected to hydrogen ion replacement treatment. Subsequently, 25 ° C. deionized water was passed through. 3000 ml of eluate was fractionated by passing the cacao crude extract and the deionized water.
Next, the 3000 ml eluate was spray-dried under a drying temperature (inlet temperature) of 160 ° C. to obtain a polyphenol-containing composition. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 59.0% by weight and the theobromine content was 0.6% by weight.
Similarly, a crude cacao extract separately prepared was freeze-dried and analyzed for components in the solid content. As a result, the polyphenol content was 28.4% by weight and the theobromine content was 8.5% by weight.
実施例5によって得たポリフェノール含有組成物を使用し、以下の評価項目から、組成物による生理効果について検討した。その結果を表1および図3に示す。 (Example 6)
Using the polyphenol-containing composition obtained in Example 5, the physiological effects of the composition were examined from the following evaluation items. The results are shown in Table 1 and FIG.
DPPH(1,1-ジフェニル-2-ピクリルヒドラジル(1,1-Dyphenyl-2-picrylhydrazyl;DPPH)ラジカル消去能は、Food Chemistry 68 (2000) Yinrong Lu, L.Yeap Foo 「Antioxidant and radical scavenging activities of polyphenols from apple pomace」に記載の方法に従い、市販のエピカテキンを指標として測定した。測定方法の概略は、以下の通りである。
指標のエピカテキンおよび各サンプルを50重量%エタノール水溶液にて溶解した。この溶液100μlに0.1mM DPPH溶液2mlを加え、30分間室温で静置した後、520nmの吸光度を測定した。ブランク溶液(DPPH溶液)の吸光度の値を阻害率100%として、各サンプルの阻害率を算出した。測定で得られた数値が低いほど、DPPHラジカル消去能が高いことを示す。 (DPPH radical scavenging ability)
DPPH (1,1-Dyphenyl-2-picrylhydrazyl; DPPH) radical scavenging ability is reported in Food Chemistry 68 (2000) Yinrong Lu, L. Yeap Foo “Antioxidant and radical scavenging According to the method described in "Activities of polyphenols from apple pomace", measurement was performed using commercially available epicatechin as an index.
The index epicatechin and each sample were dissolved in 50 wt% aqueous ethanol solution. To 100 μl of this solution, 2 ml of 0.1 mM DPPH solution was added and allowed to stand at room temperature for 30 minutes, and then the absorbance at 520 nm was measured. The inhibition rate of each sample was calculated using the absorbance value of the blank solution (DPPH solution) as the inhibition rate of 100%. It shows that DPPH radical scavenging ability is so high that the numerical value obtained by the measurement is low.
ミセル不溶化能は、Journal of Nutrition ,1999,129:1725-1730 Satoshi
Nagaoka, Kenji Miwa, Michiko Eto, Yasuo Kuzuya,Goro Hori and Kazuhiro Yamamoto「Soy Protein Peptic Hydrolysate with Bound Phospholipids Decreases Micellar Solubility and Cholesterol Absorption in Rats and Caco-2 Cells」に記載の方法に従い、市販のエピカテキンを指標として測定した。
測定方法の概略は、以下の通りである。タウロコール酸ナトリウム 7.44mmol/L、コレステロール 1.94mmol/L、オレイン酸 1mmol/L、モノオレイン酸 0.5mmol/L、フォスファチジルコリン 0.6mmol/L、塩化ナトリウム 132mmol/Lをリン酸Buffer(pH7.4)に分散させ、10分間超音波処理をしてミセル溶液を調整した。この溶液に、指標のエピカテキン及びサンプルを50重量%エタノール水溶液と良く混合し、37℃で60分間加温した。その後、遠心分離にて沈殿物を取り除き、上清液のコレステロール量を市販のコレステロールE-テストワコー(和光純薬工業株式会社製)のキットを用いて測定した。測定で得られた数値が高いほど、ミセル不溶化能が高いことを示す。 (Micellar insolubility)
Micellar insolubilization ability is described in Journal of Nutrition, 1999,129: 1725-1730 Satoshi
Nagaoka, Kenji Miwa, Michiko Eto, Yasuo Kuzuya, Goro Hori, and Kazuhiro Yamamoto As measured.
The outline of the measuring method is as follows. Sodium taurocholate 7.44 mmol / L, cholesterol 1.94 mmol / L, oleic acid 1 mmol / L, monooleic acid 0.5 mmol / L, phosphatidylcholine 0.6 mmol / L, sodium chloride 132 mmol / L The mixture was dispersed in (pH 7.4) and sonicated for 10 minutes to prepare a micelle solution. The index epicatechin and the sample were mixed well with a 50 wt% aqueous ethanol solution and heated at 37 ° C. for 60 minutes. Thereafter, the precipitate was removed by centrifugation, and the amount of cholesterol in the supernatant was measured using a commercially available kit of cholesterol E-Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The higher the numerical value obtained by measurement, the higher the micelle insolubilization ability.
市販品のカカオポリフェノール素材(株式会社明治フードマテリア製、商品名「カカオポリフェノール」)を使用し、実施例6と同様の検討を行った。その結果を表1および図3に示す。 (Comparative Example 1)
A commercially available cocoa polyphenol material (manufactured by Meiji Food Materia Co., Ltd., trade name “cocoa polyphenol”) was used, and the same examination as in Example 6 was performed. The results are shown in Table 1 and FIG.
本実施例は、分離精製の工程で使用する陽イオン交換樹脂の検討に関する。カカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.0重量%、テオブロミン含量1.2重量%)を使用し、以下の手順にしたがって、ポリフェノール含有組成物を調製した。
先ず、カカオ豆10gを磨砕した後、50重量%エタノール水溶液を200ml添加し、50℃にて30分間撹拌し、次いで、不溶成分を遠心分離処理にて除去してカカオ粗抽出液を得た。
次に、予め水素イオン置換処理した各種陽イオン交換樹脂(表2を参照)200mlを充填したカラムに、カカオ抽出液をSV=5の流速で通液した。続いて、カラムに25℃の脱イオン水を通液した。上記カカオ粗抽出液および上記脱イオン水の通液によって、溶出液600mlを分取した。
上記溶出液600mlを凍結乾燥することによりポリフェノール含有組成物を得た。得られた組成物の固形分中の成分を分析し、各組成物における成分を比較することによって、樹脂の種類による影響について検討した。その結果を表2に示す。 (Example 7)
This example relates to examination of a cation exchange resin used in the separation and purification process. A cocoa bean (dried in the production area, cocoa polyphenol content 7.0% by weight, theobromine content 1.2% by weight) was used, and a polyphenol-containing composition was prepared according to the following procedure.
First, after crushing 10 g of cocoa beans, 200 ml of 50 wt% ethanol aqueous solution was added and stirred at 50 ° C. for 30 minutes, and then insoluble components were removed by centrifugation to obtain a cacao crude extract. .
Next, the cacao extract was passed at a flow rate of SV = 5 through a column packed with 200 ml of various cation exchange resins (see Table 2) that had been previously subjected to hydrogen ion replacement treatment. Subsequently, 25 ° C. deionized water was passed through the column. 600 ml of eluate was fractionated by passing the cacao crude extract and the deionized water.
A polyphenol-containing composition was obtained by freeze-drying 600 ml of the eluate. By analyzing the components in the solid content of the obtained composition and comparing the components in each composition, the effect of the type of resin was examined. The results are shown in Table 2.
実施例7において使用した各種陽イオン交換樹脂にかえて、水酸化物イオン置換処理した陰イオン交換樹脂(表2を参照)を使用したことを除き、実施例7と同様の操作を行うことによって、溶出液を得た。次に、実施例と同様にして溶出液を凍結乾燥することによって、ポリフェノール含有組成物を得た。得られた組成物の固形分中の成分について、実施例7と同様にして分析を行った。その結果を表2に示す。 (Comparative Example 2)
By performing the same operation as in Example 7, except that instead of the various cation exchange resins used in Example 7, an anion exchange resin (see Table 2) subjected to hydroxide ion substitution treatment was used. The eluate was obtained. Next, the eluate was freeze-dried in the same manner as in Example to obtain a polyphenol-containing composition. The components in the solid content of the obtained composition were analyzed in the same manner as in Example 7. The results are shown in Table 2.
(1)各含量は、固形分の全重量を基準としたときの含有率である。
(2)「ダイヤイオン」(登録商標)のシリーズは、三菱化学株式会社製である。
(3)「アンバーライト」(登録商標)のシリーズは、ロームアンドハース株式会社製である。 Note:
(1) Each content is a content rate based on the total weight of the solid content.
(2) The “Diaion” (registered trademark) series is manufactured by Mitsubishi Chemical Corporation.
(3) A series of “Amberlite” (registered trademark) is manufactured by Rohm and Haas Co., Ltd.
本実施例は、陽イオン交換樹脂を用いた分離精製の工程で使用する溶媒の検討に関する。具体的には、陽イオン交換樹脂を充填したカラムの平衡化溶液、およびカラムからの溶離液として、種々の溶媒を使用して検討を行った。
カカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.4重量%、テオブロミン含量1.3重量%)を使用し、以下の手順にしたがって、ポリフェノール含有組成物を調製した。
先ず、カカオ豆2gを磨砕した後、50重量%エタノール水溶液を20ml添加し、50℃にて30分間撹拌し、次いで、不溶成分を遠心分離処理にて除去してカカオ粗抽出液を得た。
次に、予め、水素イオン置換処理した陽イオン交換樹脂(アンバーライト IR-120B 強酸性陽イオン交換樹脂)を20ml充填したカラムに、下記表3に示す各種平衡化溶液を100ml通液した。その後、このカカオ粗抽出液をSV=5で通液した。続いて、上記カラムに表3に示すカラム溶離液を通液した。
次に、カカオ粗抽出液およびカラム溶離液の通液によって、各溶出液を分取し、それぞれ凍結乾燥することによって、各ポリフェノール含有組成物を得た。各組成物の固形分の成分についてそれぞれ分析し、その成分を比較することによって、溶媒の影響について検討した。その結果を表3に示す。 (Example 8)
This example relates to the examination of the solvent used in the process of separation and purification using a cation exchange resin. Specifically, various types of solvents were used as the equilibration solution for the column packed with the cation exchange resin and the eluent from the column.
A cocoa bean (dried in the production area, cocoa polyphenol content 7.4% by weight, theobromine content 1.3% by weight) was used, and a polyphenol-containing composition was prepared according to the following procedure.
First, after grinding 2 g of cocoa beans, 20 ml of 50 wt% aqueous ethanol solution was added, stirred at 50 ° C. for 30 minutes, and then insoluble components were removed by centrifugation to obtain a crude cocoa extract. .
Next, 100 ml of various equilibration solutions shown in Table 3 below were passed through a column packed with 20 ml of a cation exchange resin (Amberlite IR-120B strongly acidic cation exchange resin) that had been previously subjected to hydrogen ion substitution treatment. Then, this cacao crude extract was passed at SV = 5. Subsequently, the column eluent shown in Table 3 was passed through the column.
Next, each eluate was collected by passing through a cacao crude extract and a column eluent, and lyophilized to obtain each polyphenol-containing composition. The components of the solid content of each composition were analyzed, and the effects of the solvent were examined by comparing the components. The results are shown in Table 3.
(1)平衡化溶液の温度は、それぞれ25℃である。
(2)カラム溶離液の温度は、それぞれ25℃である。
(3)各含量は、固形分の全重量を基準としたときの含有率である。 Note:
(1) The temperature of the equilibration solution is 25 ° C.
(2) The temperature of the column eluent is 25 ° C.
(3) Each content is a content rate based on the total weight of the solid content.
カカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.3重量%、テオブロミン含量1.3重量%)を使用し、以下の手順にしたがって、ポリフェノール含有組成物を調製した。
先ず、カカオ豆100gを磨砕した後、50重量%エタノール水溶液を1000ml添加し、50℃にて30分間撹拌し、次いで、不溶成分を遠心分離処理にて除去することにより、カカオ粗抽出液を得た。
次に、予め、水素イオン置換処理した陽イオン交換樹脂(アンバーライトIR-120B 強酸性陽イオン交換樹脂)を20ml充填したカラムに、上記カカオ粗抽出液をSV=5の流速で通液した。続いて、カラムに25℃の脱イオン水を通液した。上記カカオ粗抽出液および脱イオン水の通液によって、溶出液600mlを分取した。
次に、上記溶出液600mlを減圧濃縮することによって40mlの濃縮液を得た。この濃縮液を18時間静置した後、析出した沈殿を遠心分離によって、取り除いた。
次に、先の操作で得た遠心上清液を、再度、予め水素イオン置換処理した陽イオン交換樹脂(アンバーライト IR-120B)を80ml充填したカラムに、SV=5の流速で通液した。続いて、25℃の脱イオン水を通液した。上記遠心上清液および上記脱イオン水を通液によって、溶出液200mlを分取した。
次に、上記溶出液200mlを凍結乾燥することにより、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は52.9%、テオブロミン含量は0.9%であった。 Example 9
A cocoa bean (dried in the production area, cocoa polyphenol content 7.3% by weight, theobromine content 1.3% by weight) was used, and a polyphenol-containing composition was prepared according to the following procedure.
First, after grinding 100 g of cocoa beans, 1000 ml of a 50 wt% aqueous ethanol solution was added, stirred at 50 ° C. for 30 minutes, and then the insoluble components were removed by centrifugal separation treatment to obtain a crude cocoa extract. Obtained.
Next, the crude cacao extract was passed through the column filled with 20 ml of a cation exchange resin (Amberlite IR-120B strong acid cation exchange resin) that had been previously subjected to hydrogen ion substitution treatment at a flow rate of SV = 5. Subsequently, 25 ° C. deionized water was passed through the column. By passing the above cacao crude extract and deionized water, 600 ml of the eluate was collected.
Next, the eluate (600 ml) was concentrated under reduced pressure to obtain a 40 ml concentrate. The concentrated solution was allowed to stand for 18 hours, and then the deposited precipitate was removed by centrifugation.
Next, the centrifugation supernatant obtained in the previous operation was again passed through a column packed with 80 ml of a cation exchange resin (Amberlite IR-120B) that had been previously subjected to hydrogen ion substitution at a flow rate of SV = 5. . Subsequently, 25 ° C. deionized water was passed through. 200 ml of the eluate was collected by passing through the centrifugal supernatant and the deionized water.
Next, 200 ml of the eluate was freeze-dried to obtain a polyphenol-containing composition. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 52.9% and the theobromine content was 0.9%.
カカオ豆(産地で乾燥を終えたもの、カカオポリフェノール含量7.3重量%、テオブロミン含量1.3重量%)を使用し、以下の手順にしたがって、ポリフェノール含有組成物を調製した。
先ず、カカオ豆100gを磨砕した後、50重量%エタノール水溶液を1000ml添加し、50℃にて30分間撹拌し、次いで、不溶成分を遠心分離処理にて除去することによってカカオ粗抽出液を得た。
次に、予め水素イオン置換処理した陽イオン交換樹脂(アンバーライトIR-120B 強酸性陽イオン交換樹脂)を20ml充填したカラムに、上記カカオ粗抽出液をSV=5の流速で通液した。続いて、カラムに25℃の脱イオン水を通液した。上記カカオ粗抽出液および上記脱イオン水の通液によって、溶出液600mlを分取した。引き続き、上記溶出液600mlを減圧濃縮することによって40mlの濃縮液を得た。
次に、先の操作で得た濃縮液を、再度、予め水素イオン置換処理した陽イオン交換樹脂(アンバーライト IR-120B)を160ml充填したカラムに、SV=5の流速で通液した。続いて、25℃の脱イオン水を通液した。上記濃縮液および上記脱イオン水の通液によって、溶出液360mlを分取した。 次に、上記溶出液360mlを凍結乾燥することにより、ポリフェノール含有組成物を得た。この組成物の固形分中の成分を分析した結果、ポリフェノール含量は51.9%、テオブロミン含量は0.9%であった。 (Example 10)
A cocoa bean (dried in the production area, cocoa polyphenol content 7.3% by weight, theobromine content 1.3% by weight) was used, and a polyphenol-containing composition was prepared according to the following procedure.
First, after crushing 100 g of cocoa beans, 1000 ml of 50 wt% aqueous ethanol solution was added, stirred at 50 ° C. for 30 minutes, and then insoluble components were removed by centrifugation to obtain a crude cocoa extract. It was.
Next, the cacao crude extract was passed through a column packed with 20 ml of a cation exchange resin (Amberlite IR-120B strong acid cation exchange resin) that had been previously subjected to hydrogen ion substitution treatment at a flow rate of SV = 5. Subsequently, 25 ° C. deionized water was passed through the column. 600 ml of eluate was fractionated by passing the cacao crude extract and the deionized water. Subsequently, 600 ml of the eluate was concentrated under reduced pressure to obtain 40 ml of concentrate.
Next, the concentrated solution obtained in the previous operation was passed again through a column packed with 160 ml of a cation exchange resin (Amberlite IR-120B) previously subjected to hydrogen ion substitution treatment at a flow rate of SV = 5. Subsequently, 25 ° C. deionized water was passed through. By passing the concentrated solution and the deionized water, 360 ml of the eluate was collected. Next, 360 ml of the eluate was freeze-dried to obtain a polyphenol-containing composition. As a result of analyzing the components in the solid content of this composition, the polyphenol content was 51.9% and the theobromine content was 0.9%.
Claims (10)
- 以下の工程を有するポリフェノール含有組成物の製造方法。
(a)ポリフェノールを含有する植物体またはその加工品を溶媒で抽出し、ポリフェノール粗抽出液を得る工程、
(b)前記ポリフェノール粗抽出液を、予め水素イオン置換処理した陽イオン交換樹脂に接触させ、引き続き、前記陽イオン交換樹脂に、イオン性物質を含まない第1の溶媒を通液し、第1のポリフェノール溶出液を得る工程、
(c)前記第1のポリフェノール溶出液を、予め水素イオン置換処理した陽イオン交換樹脂に再度通液した後に、イオン性物質を含まない第2の溶媒を通液して第2のポリフェノール溶出液を得る工程、および
(d)前記第2のポリフェノール溶出液を濃縮または乾燥させる工程。 The manufacturing method of the polyphenol containing composition which has the following processes.
(A) extracting a plant body containing polyphenol or a processed product thereof with a solvent to obtain a polyphenol crude extract,
(B) The polyphenol crude extract is brought into contact with a cation exchange resin that has been subjected to a hydrogen ion replacement treatment in advance, and subsequently, a first solvent that does not contain an ionic substance is passed through the cation exchange resin. Obtaining a polyphenol eluate of
(C) The first polyphenol eluate is again passed through a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then a second solvent that does not contain an ionic substance is passed through to pass through the second polyphenol eluate. And (d) concentrating or drying the second polyphenol eluate. - 前記植物体がカカオであることを特徴とする、請求項1に記載の製造方法。 The production method according to claim 1, wherein the plant body is cacao.
- 前記工程(a)において、前記抽出に用いる溶媒が、40~70重量%のエタノール水溶液である、請求項1または2に記載の製造方法。 The production method according to claim 1 or 2, wherein in the step (a), the solvent used for the extraction is a 40 to 70 wt% aqueous ethanol solution.
- 前記工程(b)及び前記工程(c)において、前記第1および第2の溶媒の通液がSV=2~10の流速で実施されることを特徴とする、請求項1~3のいずれかに記載の製造方法。 The process according to any one of claims 1 to 3, wherein in the step (b) and the step (c), the first and second solvents are passed at a flow rate of SV = 2 to 10. The manufacturing method as described in.
- 前記工程(c)に先立ち、前記第1のポリフェノール溶出液からテオブロミンの沈殿物を析出させ、次いで前記沈殿物を除去することによって得られる溶液を前記工程(c)で使用することを特徴とする、請求項1~4のいずれかに記載の製造方法。 Prior to the step (c), a solution obtained by precipitating a theobromine precipitate from the first polyphenol eluate and then removing the precipitate is used in the step (c). The production method according to any one of claims 1 to 4.
- 前記イオン性物質を含まない第1および第2の溶媒が、それぞれ、脱イオン水またはエタノール水溶液である、請求項1~5のいずれかに記載の製造方法。 The production method according to any one of claims 1 to 5, wherein the first and second solvents not containing the ionic substance are each deionized water or an aqueous ethanol solution.
- 前記イオン性物質を含まない第2の溶媒が、35℃以下の脱イオン水またはエタノール濃度が30重量%以下のエタノール水溶液である、請求項1~6のいずれかに記載の製造方法。 The production method according to any one of claims 1 to 6, wherein the second solvent not containing the ionic substance is deionized water of 35 ° C or lower or an aqueous ethanol solution having an ethanol concentration of 30 wt% or lower.
- 請求項1~7に記載の製造方法によって得られるポリフェノール含有組成物。 A polyphenol-containing composition obtained by the production method according to claims 1 to 7.
- 固形分中のポリフェノール含量が33重量%以上であり、テオブロミン含量が1重量%以下であるポリフェノール含有組成物。 A polyphenol-containing composition having a polyphenol content in a solid content of 33% by weight or more and a theobromine content of 1% by weight or less.
- ポリフェノールを含有する植物体またはその加工品からポリフェノール含有組成物を製造するための装置であって、
ポリフェノールを含有する植物体またはその加工品からポリフェノール粗抽出液を得る手段を有する抽出区分と、
前記抽出区分から移送手段を介して移送されたポリフェノール粗抽出液を、予め水素イオン置換処理した陽イオン交換樹脂に接触させ、引き続き上記陽イオン交換樹脂に、イオン性物質を含まない第1の溶媒を通液し、第1のポリフェノール溶出液を得る手段を有する第1の分離精製、および前記第1の分離精製区分から移送手段を介して移送された上記第1のポリフェノール溶出液を、予め水素イオン置換処理した陽イオン交換樹脂に再度通液した後に、イオン性物質を含まない第2の溶媒を通液して第2のポリフェノール溶出液を得る手段を有する第2の分離精製を可能とする分離精製区分と、
前記分離精製区分から移送手段を介して移送された上記第2のポリフェノール溶出液を濃縮または乾燥する手段を有する回収区分と
を有することを特徴とする製造装置。 An apparatus for producing a polyphenol-containing composition from a plant containing polyphenol or a processed product thereof,
An extraction section having means for obtaining a polyphenol crude extract from a plant containing polyphenol or a processed product thereof;
The crude polyphenol extract transferred from the extraction section via the transfer means is brought into contact with a cation exchange resin that has been previously subjected to hydrogen ion substitution treatment, and then the first solvent that does not contain an ionic substance in the cation exchange resin. The first polyphenol eluate having a means for obtaining a first polyphenol eluate through which the liquid is passed, and the first polyphenol eluate transferred from the first separation / purification section via the transfer means are previously hydrogenated. After passing again through the ion-exchange-treated cation exchange resin, the second separation / purification having means for obtaining a second polyphenol eluate by passing a second solvent not containing an ionic substance is enabled. Separation and purification division,
And a recovery section having a means for concentrating or drying the second polyphenol eluate transferred from the separation and purification section through a transfer means.
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JPH11228565A (en) * | 1998-02-10 | 1999-08-24 | T Hasegawa Co Ltd | Tea leaf tannin, its production, and drink and food containing the same |
JPH11292870A (en) * | 1998-04-02 | 1999-10-26 | T Hasegawa Co Ltd | Production of tea leaf tannins |
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JPWO2009099221A1 (en) * | 2008-02-07 | 2011-06-02 | 明治製菓株式会社 | Method and apparatus for producing a theobromine-rich composition |
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