JP2017046601A - Manufacturing method of purified catechins-containing composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a manufacturing method of a purified catechins-containing composition capable of recovering catechins at good purity and high yield.SOLUTION: There is provided a manufacturing method of a purified catechins-containing composition including a process A of mixing a catechins-containing composition having a mass ratio of (A) catechins with content of 1 to 35 mass% and (B) phospholipid [(B)/(A)] of 2 to 100 and at least one kind of solvent selected from an organic solvent solution and an organic solvent and a process B of recovering a phase rich in catechins generated in a mixture. There is provided a manufacturing method of a purified catechins-containing composition, where the organic solvent is preferably ethanol, organic solvent concentration is 15 to 55 mass. There is provided a manufacturing method of a purified catechins-containing composition, where the phase rich in catechins is a liquid phase obtained by solid solution separation of the mixture.SELECTED DRAWING: None

Description

  The present invention relates to a method for producing a purified catechins-containing composition.

  Tea leaves are rich in catechins, and catechins can be obtained by extraction from tea leaves, but impurities such as caffeine are also extracted at the time of extraction.

  As a technique for reducing such impurities, for example, tea leaves are extracted with hot water or an organic solvent aqueous solution, a solution containing the extracted components is washed with chloroform, and then the extracted components are transferred into an organic solvent, and then the organic solvent is added. Method of distilling (Patent Document 1), method of bringing tea extract into contact with activated clay or acidic clay (Patent Document 2), dissolving or suspending tea extract in water or water-containing organic solvent, synthesis under alkaline conditions A method of contacting with an adsorbent (Patent Document 3) is known.

  On the other hand, it has been reported that the astringency of tea extract can be reduced by coexisting tea extract with phospholipid (Patent Document 4), but a method for purifying tea extract using phospholipid is known. Not.

JP 59-219384 A JP-A-6-142405 JP-A-8-109178 Special table 2012-503479 gazette

  An object of the present invention is to provide a method for producing a purified catechins-containing composition capable of recovering catechins with high purity and higher yield.

  The inventor mixed a catechins-containing composition containing catechins and phospholipids in a specific ratio with a specific solvent, so that a phase rich in catechins and a phase rich in phospholipids and contaminants were obtained. It was found that catechins can be obtained with high purity and higher yield by recovering the separated and rich phase of catechins.

  That is, the present invention relates to a catechin-containing composition in which the mass ratio [(B) / (A))] of (A) catechins to (B) phospholipids is 2 to 100, an organic solvent aqueous solution, and an organic solvent A method for producing a purified catechins-containing composition comprising: a step A in which at least one solvent selected from the above is mixed; and a step B in which a phase rich in catechins generated in the mixed solution is recovered. .

  ADVANTAGE OF THE INVENTION According to this invention, the manufacturing method of the refined catechins containing composition which can collect | recover catechins with sufficient purity and a higher yield can be provided.

  The method for producing a purified catechins-containing composition of the present invention includes Step A and Step B. Hereinafter, each step will be described.

<Process A>
Step A is selected from a catechin-containing composition having a mass ratio [(B) / (A)] of (A) catechins to (B) phospholipids of 2 to 100, an organic solvent aqueous solution, and an organic solvent. And at least one solvent.

Here, in the present specification, “catechins” refers to a gallate body composed of epigallocatechin gallate, gallocatechin gallate, epicatechin gallate and catechin gallate, and a non-gallate body composed of epigallocatechin, gallocatechin, epicatechin and catechin. Is a collective term. The content of catechins is defined based on the total amount of the above eight types, and in the present invention, at least one of the above eight types of catechins may be contained.
In the present specification, “phospholipid” is a generic term for lipids having a phosphate ester moiety and a phosphonate ester moiety in the structure.

(Catechin-containing composition)
The catechins-containing composition includes (A) catechins and (B) phospholipids, and their mass ratio [(B) / (A))] is 2 to 100. From the viewpoint of rate and purity, 2 or more is preferable, 4 or more is more preferable, 6 or more is more preferable, 100 or less is preferable, 50 or less is more preferable, 25 or less is further preferable, and 20 or less is even more preferable. As a range of this mass ratio, 2-100 are preferable, 4-50 are more preferable, 6-25 are still more preferable, and 6-20 are still more preferable.

  From the viewpoint of the yield and purity of catechins, the content of (A) catechins in the solid content of the catechin-containing composition is preferably 1% by mass or more, more preferably 2% by mass or more, and 4% by mass or more. Is more preferably 35% by mass or less, more preferably 30% by mass or less, and further preferably 25% by mass or less. The range of the content of the (A) catechins is preferably 1 to 35% by mass, more preferably 2 to 30% by mass, and further preferably 4 to 25% by mass in the solid content of the catechins-containing composition. It is. Here, the “solid content” in this specification is a residue obtained by drying a sample for 6 hours with an electric constant temperature dryer at 105 ° C. to remove volatile substances, and is calculated by the following formula (I).

  Solid content (mass%) = mass after sample drying (g) / mass before sample drying (g) × 100 (I)

  The content of (B) phospholipid in the solid content in the catechin-containing composition is preferably 65% by mass or more, more preferably 70% by mass or more, and more preferably 75% by mass or more from the viewpoint of ease of separation operation. Further, from the viewpoint of the yield and purity of catechins, it is preferably 99% by mass or less, more preferably 98% by mass or less, and still more preferably 96% by mass or less. The content range of the (B) phospholipid is preferably 65 to 99% by mass, more preferably 70 to 98% by mass, and still more preferably 75 to 96% by mass in the solid content in the catechins-containing composition. %. The amount of phospholipid in the catechin-containing composition can be quantified by, for example, a phospholipid quantification kit such as phospholipid C test Wako (manufactured by Wako Pure Chemical Industries), analysis using HPLC, or the like. .

  In the catechin-containing composition, the mass ratio [(C) / (A)] of (A) catechins to (C) caffeine is preferably 1 or less, more preferably 0.5 or less, and 0.1 or less. Is more preferable, and 0.05 or less is even more preferable. The lower limit is not particularly limited and may be 0. However, for example, 0.00001 or more is preferable, and 0.0001 or more is more preferable. The range of the mass ratio [(C) / (A)] is preferably 0.00001 to 1, more preferably 0.00001 to 0.5, still more preferably 0.00001 to 0.1, and still more preferably. Is 0.0001-0.05.

(One aspect for preparing a catechin-containing composition)
The catechins-containing composition may be any composition as long as the mass ratio [(B) / (A)] of (A) catechins and (B) phospholipid is 2 to 100, preferably satisfies the above-mentioned preferred range, Although it can be prepared by an appropriate method, for example, it is prepared by (A ′) mixing a tea extract and a phospholipid, and subjecting the mixture to a method including a step of recovering the phospholipid phase from the mixed solution. be able to.

(A ′) Examples of the tea extract include a tea extract, a concentrate thereof, or a purified product thereof, and there are various forms such as a solid, a liquid, a solution, and a slurry. The tea extract can be used alone or in combination of two or more. As used herein, “tea extract” refers to a product extracted from tea leaves using hot water or a hydrophilic organic solvent, and has not been concentrated or purified. The extraction method and extraction conditions can employ known methods and conditions, and are not particularly limited. Examples of tea leaves include the genus Camellia, for example, C. sinensis var. Sinensis (including Yabuta species), C. sinensis var. Assamica, and tea trees (Camellia sinensis) selected from hybrids thereof. Tea trees can be classified into non-fermented tea, semi-fermented tea, and fermented tea according to their processing methods.
Examples of non-fermented tea include green tea such as sencha, bancha, mochi tea, kettle tea, stem tea, stick tea, and bud tea. Examples of the semi-fermented tea include oolong tea such as iron kannon, color type, golden katsura, and martial arts tea. Further, examples of fermented tea include black teas such as Darjeeling, Assam, Sri Lanka and the like. These can be used alone or in combination of two or more. Among these, non-fermented tea is preferable and green tea is more preferable from the viewpoint of the content of catechins.

  The “tea extract concentrate” is a product obtained by removing a part of the solvent from the tea extract to increase the concentration of catechins. For example, as a concentration method, atmospheric concentration, reduced pressure concentration, membrane concentration Etc. Commercially available products may be used as the concentrate of the tea extract. For example, green tea such as “Polyphenone” from Mitsui Norin Co., “Theafranc” from ITO EN, “Sunphenon” from Taiyo Kagaku Co., Ltd. A concentrate of the extract may be mentioned. Furthermore, the “purified product of tea extract” is a product obtained by purifying a tea extract or a concentrate thereof to increase the purity of catechins. For example, JP 2004-147508 A and JP 2004-149416 A The methods described in Japanese Patent Laid-Open No. 2006-160656, Japanese Patent Laid-Open No. 2007-282568, Japanese Patent Laid-Open No. 2008-079609, and the like can be employed. In addition, a product obtained by subjecting the tea extract to various chromatographies such as reverse phase chromatography and liquid chromatography to fractionate catechins may be used.

  The tea extract is preferably in the form of an aqueous solution. The aqueous solution in which the tea extract is dissolved can be obtained by, for example, diluting or concentrating the tea extract extracted from tea leaves using water, if necessary. Even if it dilutes and uses it, you may use again after dissolving the tea extract, its concentrate, or those dried dried products in water.

  The content of catechins in the aqueous solution in which the tea extract is dissolved can be selected as appropriate. However, from the viewpoint of purification efficiency, 0.02% by mass or more is preferable, 0.05% by mass or more is more preferable, and 06 mass% or more is still more preferable, 10 mass% or less is preferable, 7 mass% or less is more preferable, and 5 mass% or less is still more preferable. As a range of content of this catechin, Preferably it is 0.02-10 mass%, More preferably, it is 0.05-7 mass%, More preferably, it is 0.06-5 mass%.

  In addition, the tea extract used in the present invention has a gallate body ratio (hereinafter also referred to as “gallate body ratio”) in catechins of preferably 30% by mass or more and 35% by mass or more from the viewpoint of physiological effects. More preferably, 40% by mass or more is further preferable, and from the viewpoint of flavor, 70% by mass or less is preferable, 65% by mass or less is more preferable, and 60% by mass or less is more preferable. The range of the gallate body ratio is preferably 30 to 70% by mass, more preferably 35 to 65% by mass, and still more preferably 40 to 60% by mass. Here, in this specification, the “gallate body ratio” is a mass ratio of the four gallate bodies to the eight catechins.

  (B) As for phospholipid, those derived from egg yolk, soybean and other animal and plant materials can be used without particular limitation, and semi-synthetic phospholipids such as hydrogenated products and hydroxide derivatives thereof, synthetic products Etc. (B) The constituent fatty acid of the phospholipid is not particularly limited, and may be either a saturated fatty acid or an unsaturated fatty acid. The (B) phospholipid may contain a charged phospholipid such as an anionic phospholipid and a cationic phospholipid, and further a polymerizable phospholipid in addition to the neutral phospholipid. Phospholipids can be used alone or in combination of two or more.

  Examples of (B) phospholipid include glycerophospholipid, lysoglycerophospholipid, and sphingophospholipid. Of these, glycerophospholipid and sphingophospholipid are preferable. Examples of the glycerophospholipid include phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine, among which phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine are preferable. Examples of sphingophospholipids include sphingomyelin and ceramide serialine. As a raw material containing these phospholipids, natural lecithin or a purified product obtained by hydrogenating it can be used. Examples of natural lecithin include egg yolk lecithin, soybean lecithin, and squid lecithin. Examples of hydrogenated phospholipid include hydrogenated soybean phosphatidylcholine and hydrogenated egg yolk lecithin.

  The phospholipid when mixed with the tea extract may be in the form of a phospholipid membrane or a liposome. By making the phospholipid into the form of a phospholipid membrane or a liposome, the catechins can be easily taken into the phospholipid membrane or its internal aqueous phase, and the catechins can be adsorbed onto the phospholipid membrane. Here, “liposome” in the present specification refers to a closed vesicle having an inner aqueous phase part surrounded by a phospholipid bilayer membrane, and a multiphase liposome (Multilamellar Vesicle: depending on the size and the number of lipid bimolecules). MLV), large unilamellar vesicle (LUV), and small unilamellar vesicle (SUV). Any type of liposome can be used in the present invention.

When adsorbing catechins to phospholipid membranes, for example, mixing tea extract and phospholipids, applying stress by shaking, etc. to form phospholipid membranes and adsorbing non-polymer catechins to phospholipid membranes May be performed substantially simultaneously. Moreover, after applying a stress to the phospholipid by shaking or the like to form a phospholipid film in advance, the tea extract can be mixed therewith.
The method for forming the liposome is not particularly limited, and methods such as a Bangham method, a lipid dissolution method, a mechanochemical method, and a freeze-dried liposome method can be employed.

The median diameter of the liposome is preferably 10 to 20,000 nm, more preferably 30 to 15,000 nm, still more preferably 50 to 10,000 nm, and still more preferably, from the viewpoint of the separation operation of the phospholipid phase and the liquid phase. It is 50 to 5,000 nm, more preferably 50 to 3,000 nm, still more preferably 80 to 1,000 nm, and still more preferably 100 to 500 nm. As used herein, “median diameter” refers to a particle diameter corresponding to a cumulative value of 50% (d ₅₀ ) in a volume-based cumulative particle size distribution measured using a laser diffraction scattering method.

Moreover, what has a phase transition temperature for the phospholipid at the time of mixing with a tea extract is preferable. Here, in this specification, the “phase transition temperature” means a temperature at which a phase transition between both states of a gel and a liquid crystal that can be taken by the phospholipid occurs, and when there is a sufficient amount of water relative to the phospholipid. Is the value of The phase transition temperature can be measured by differential thermal analysis using a differential scanning calorimeter (DSC).
For example, DSC7020 (manufactured by Hitachi High-Tech Science) can be used as the differential scanning calorimeter. Specifically, the phase transition temperature of phospholipid can be measured by the following procedure.
(1) A phospholipid having a phase transition temperature at a temperature exceeding 0 ° C. measures the phase transition temperature according to the following procedure. First, a sample is prepared in which phospholipid is sufficiently dispersed in water so that the phospholipid concentration is 1 to 10 g / 100 g so that a sufficient amount of water is present relative to the phospholipid. Next, 0.005 g of a sample is weighed into an aluminum sample container, covered with an aluminum, and then sealed using an electric sample sealer. Place the sealed sample container on the holder unit in the electric furnace. Also, a blank container sealed with air is placed on the blank side of the holder unit. Cover the electric furnace and hold at 1 ° C. for 5 minutes in a nitrogen atmosphere. Subsequently, it heats from 1 degreeC to 98 degreeC with the temperature increase rate of 0.5 degreeC / min (temperature rising process). In this temperature raising step, a DSC curve is measured using a differential scanning calorimeter DSC7020 (manufactured by Hitachi High-Tech Science). At this time, the temperature when the endothermic peak is observed in the temperature raising step is defined as the phase transition temperature of the phospholipid.
(2) For phospholipids whose phase transition temperature cannot be confirmed by the above measurement conditions, the phase transition temperature is measured by the following procedure. First, a sample in which a 50% by mass ethylene glycol aqueous solution with respect to phospholipid is sufficiently dispersed in water so as to have a phospholipid concentration of 1 to 10 g / 100 g is prepared. Next, a sample container and a blank container are prepared by the same operation as described above, and placed on the holder unit. Cover the electric furnace and hold at 25 ° C. for 5 minutes under a nitrogen atmosphere. Subsequently, it cools from 25 degreeC to -40 degreeC with the temperature-fall rate of 0.5 degree-C / min (cooling process). In this cooling step, a DSC curve is measured using a differential scanning calorimeter DSC7020 (manufactured by Hitachi High-Tech Science). At this time, the temperature at which an exothermic peak is observed in the cooling step is defined as the phase transition temperature of the phospholipid.

  The phase transition temperature of the phospholipid is preferably −25 ° C. or higher, more preferably −20 ° C. or higher, from the viewpoint of handling, and preferably 60 ° C. or lower, and 50 ° C. or lower, from the viewpoint of catechin uptake and adsorption rate. Is more preferable, 40 degrees C or less is still more preferable, and 30 degrees C or less is still more preferable. The range of the phase transition temperature is preferably -25 ° C to 60 ° C, more preferably -20 ° C to 50 ° C, still more preferably -20 ° C to 40 ° C, still more preferably -20 ° C to 30 ° C. .

Examples of phospholipids having a phase transition temperature include soybean phosphatidylcholine (phase transition temperature measured by the measurement conditions in the present specification is about -10 ° C), hydrogenated soybean phosphatidylcholine (phase transition temperature measured by the measurement conditions in the present specification). About 51 ° C.), dimyristoyl phosphatidylcholine (phase transition temperature described in Reference 1 below about 23 ° C.), dipalmitoyl phosphatidylcholine (phase transition temperature described below Reference 1 about 41 ° C.), distearoyl phosphatidylcholine (reference below) Phase transition temperature described in Reference 1 is about 54 ° C., dihexadecyl phosphatidyl choline (phase transition temperature described in Reference 2 below is about 45 ° C.), stearoyl palmitol phosphatidyl choline (phase transition temperature described in Reference 3 below is about 45). ℃), dipalmitoylphosphatidyl ester Noruamin (Reference 2 phase transition temperature of about 63 ° C. of the following description), and the like. Of these, phosphatidylcholine having a choline group is preferable. As the phospholipid used in the present invention, a commercially available product or a synthetic product can be appropriately selected and used. The phospholipid content is preferably 60% by mass or more, and 80% by mass or more. Further preferred.

Reference 1: Gold et al., Phospholipid bilayer membrane under high pressure. Science and technology of high pressure, vol9, No3, p.213-220 (1999)
Reference 2: Matsuki et al. Membrane state of biological membrane lipids-Structure-function correlations revealed from pressure studies-High pressure science and technology, vol23, No1, p.30-38 (2013)
Reference 3: Proceedings of the 32nd Physical and Physical Chemistry Meeting

  From the viewpoint of production efficiency, the mixing ratio of the tea extract and phospholipid is 0 as the mass ratio [(A) / (B)] of (A) catechins and (B) phospholipid in the tea extract. 0.001 or more is preferable, 0.01 or more is more preferable, 0.02 or more is more preferable, and from the viewpoint of the yield and purity of catechins, 0.20 or less is preferable, 0.18 or less is more preferable, and 0 .11 or less is more preferable, and 0.09 or less is particularly preferable. The range of the mass ratio [(A) / (B)] is preferably 0.001 to 0.20, more preferably 0.01 to 0.18, still more preferably 0.01 to 0.11, and particularly preferably. Preferably it is 0.02-0.09.

The mixing temperature of the tea extract and phospholipid is preferably higher than the phase transition temperature of the phospholipid used, more preferably 5 ° C. or more higher than the phase transition temperature of the phospholipid, The temperature is more preferably 10 ° C. or higher, and more preferably 20 ° C. or higher.
The mixing time of the tea extract and phospholipid is not uniform depending on the production scale and the like, but from the viewpoint of the recovery rate of catechins and purification efficiency, preferably 10 to 360 minutes, more preferably 15 to 120 minutes, further Preferably it is 20 to 60 minutes. A liquid temperature control means can be provided so that the temperature of the liquid mixture is kept constant for a predetermined time.

  The pH at the time of mixing the tea extract and phospholipid (25 ° C.) is preferably 2.5 to 9.0, more preferably 4.0 to 8 from the viewpoint of preventing equipment corrosion and the stability of catechins. 0.0, more preferably 6.0 to 7.0.

  The order of mixing the tea extract and the phospholipid is not particularly limited, and the tea extract and the phospholipid may be charged and mixed at the same time, or one may be charged and mixed into the other. Moreover, when mixing a tea extract and a phospholipid, you may perform processes, such as stirring, shaking, and ultrasonic irradiation.

  By mixing tea extract and phospholipid, catechins are taken into the phospholipid membrane or its internal aqueous phase, or adsorbed to the phospholipid membrane, while impurities are outside the phospholipid membrane or outside the liposome. Since it exists in the aqueous phase as it is, catechins and impurities are separated.

  After mixing, the phospholipid phase is recovered from the mixture, and examples of the recovery method include solid-liquid separation such as ultrafiltration, centrifugation, gel chromatography, and dialysis. Solid-liquid separation can be performed by combining one type or two or more types. Of these, centrifugation is preferred. By centrifuging the mixed solution, the phospholipid phase can be easily recovered as a precipitated phase.

As the centrifuge, general devices such as a separation plate type, a cylindrical type, and a decanter type can be used, and an ultracentrifuge may be used as necessary.
The temperature at the time of centrifugation is preferably 1 to 60 ° C., more preferably 2 to 50 ° C., and further preferably 3 to 40 ° C. from the viewpoint of the recovery rate of the phospholipid phase.
The centrifugation time is preferably 3 to 300 minutes, more preferably 5 to 200 minutes, still more preferably 10 to 100 minutes, and still more preferably 15 to 60 minutes, from the viewpoint of the recovery rate of the phospholipid phase.
In addition, the centrifugation conditions are, for example, preferably 300 G or more, more preferably 500 G or more, and more preferably 800 G or more as the relative centrifugal acceleration from the viewpoint of removing contaminants, the recovery rate of catechins, and recovering the phospholipid phase as a precipitation phase. Is more preferable. In addition, although an upper limit is not specifically limited, For example, 170000G or less is preferable. The range of the relative centrifugal acceleration is preferably 300 to 170000G, more preferably 500 to 170000G, and still more preferably 800 to 170000G. The rotation speed and the rotation radius of the centrifuge can be appropriately selected so that the relative centrifugal acceleration is within the above range. Here, “relative centrifugal acceleration” in this specification refers to a value calculated by the following equation (II).

Relative centrifugal acceleration (G) = 1188 × r × N ² × 10 ⁻⁸ (II)

[In the formula (II), r represents the maximum rotation radius (cm) of the centrifuge, and N represents the number of rotations per minute (rpm). ]

  The phospholipid phase recovered in this manner can be used as it is as the catechin-containing composition in Step A.

(solvent)
In this step, at least one selected from an organic solvent aqueous solution and an organic solvent is used as a solvent.
The organic solvent is preferably a hydrophilic organic solvent, and specific examples include alcohols such as ethanol and methanol, ketones such as acetone, and esters such as ethyl acetate. Of these, alcohols and ketones are preferable. In consideration of use in foods, alcohols are more preferable, and ethanol is more preferable.

  The organic solvent concentration in the organic solvent aqueous solution is preferably 10% by mass or more, more preferably 20% by mass or more, still more preferably 30% by mass or more, and more preferably 40% by mass or more from the viewpoint of the yield and purity of catechins. Further preferred.

(mixture)
The mixing method of the catechins-containing composition and the solvent is not particularly limited, and the catechins-containing composition and the solvent may be charged simultaneously and mixed, or one may be charged and mixed into the other. Moreover, when mixing a catechins containing composition and a solvent, you may perform processes, such as stirring, shaking, and ultrasonic irradiation.

  From the viewpoint of the yield and purity of catechins, the amount of solvent used is preferably 2 or more, more preferably 4 or more, and more preferably 6 or more as the mass ratio of the total amount of solvent to the total solid content in the catechins-containing composition. Further, from the viewpoint of productivity, 70 or less is preferable, 60 or less is more preferable, and 50 or less is more preferable. The range of the mass ratio is preferably 2 to 70, more preferably 4 to 60, and further preferably 6 to 50. The “total solvent amount” referred to here is the sum of the amount of the solvent (for example, water) contained in the catechins-containing composition and the amount of the organic solvent aqueous solution or the organic solvent used for mixing.

From the viewpoint of the yield of catechins, the organic solvent concentration in the mixed solution is preferably 15% by mass or more, more preferably 20% by mass or more, further preferably 25% by mass or more, and from the viewpoint of the purity of catechins, 55 mass% or less is preferable, 53 mass% or less is more preferable, and 50 mass% or less is still more preferable. The range of the organic solvent concentration is preferably 15 to 55% by mass, more preferably 20 to 53% by mass, and still more preferably 25 to 50% by mass.
As a method for adjusting the concentration of the organic solvent in the mixed solution, for example, a method of mixing the catechins-containing composition and the organic solvent aqueous solution to adjust the organic solvent concentration within the above range, the catechins-containing composition is water. After dissolution, the organic solvent is added to adjust the organic solvent concentration within the above range. After the catechin-containing composition is dispersed or dissolved in the organic solvent, water is gradually added to keep the organic solvent concentration within the above range. The method of adjusting to, etc. are mentioned. The catechin-containing composition may contain a solvent. When the catechin-containing composition contains a solvent, the amount of the organic solvent and / or water added in consideration of the concentration of the solvent. Is appropriately determined, and the concentration of the organic solvent in the mixed solution may be adjusted.

The mixing temperature of the catechins-containing composition and the solvent is preferably 5 ° C. or higher, more preferably 10 ° C. or higher, more preferably 20 ° C. or higher, even more preferably 30 ° C. or higher, from the viewpoint of the recovery rate and purity of catechins. Moreover, from a stability viewpoint of catechins, 60 degrees C or less is preferable, 55 degrees C or less is more preferable, and 50 degrees C or less is still more preferable. The range of the mixing temperature is preferably 5 to 60 ° C, more preferably 10 to 55 ° C, still more preferably 20 to 55 ° C, and still more preferably 30 to 50 ° C.
The mixing time of the catechin-containing composition and the solvent is not uniform depending on the production scale and the like, but from the viewpoint of the recovery rate and purity of catechins, purification efficiency, preferably 10 minutes or more, more preferably 15 minutes or more, 20 minutes or more is more preferable, and 360 minutes or less is preferable, 120 minutes or less is more preferable, and 60 minutes or less is still more preferable. The range of the mixing time is preferably 10 to 360 minutes, more preferably 15 to 120 minutes, and further preferably 20 to 60 minutes. A liquid temperature control means can be provided so that the temperature of the liquid mixture is kept constant for a predetermined time.

<Process B>
Step B is a step of recovering a phase rich in catechins generated in the mixed solution obtained in Step A.
In this step, the liquid phase produced by solid-liquid separation of the mixed solution can be recovered as a phase rich in catechins. Examples of the solid-liquid separation include ultrafiltration, centrifugation, gel chromatography, dialysis and the like, as described above. Solid-liquid separation can be performed by combining one type or two or more types. Of these, centrifugation is preferred. By centrifuging the mixed solution, a phase rich in phospholipids and contaminants is precipitated, so that a liquid phase rich in catechins dissolved in a solvent can be easily recovered as a supernatant.

As the centrifuge, general devices such as a separation plate type, a cylindrical type, and a decanter type can be used, and an ultracentrifuge may be used as necessary.
The temperature at the time of centrifugation is preferably 5 ° C. or higher, more preferably 10 ° C. or higher, further preferably 20 ° C. or higher, particularly more preferably 30 ° C. or higher, from the viewpoint of the yield of catechins and ease of separation. Moreover, from a stability viewpoint of catechins, 60 degrees C or less is preferable, 55 degrees C or less is more preferable, and 50 degrees C or less is still more preferable. The temperature range is preferably 5 to 60 ° C, more preferably 10 to 55 ° C, still more preferably 20 to 55 ° C, and still more preferably 30 to 50 ° C.
Centrifugation time is preferably 3 minutes or more, more preferably 5 minutes or more, further preferably 10 minutes or more, even more preferably 15 minutes or more, from the viewpoint of the recovery rate and purity of catechins, ease of separation, And 300 minutes or less are preferable, 200 minutes or less are more preferable, 100 minutes or less are still more preferable, and 60 minutes or less are still more preferable. The range of the centrifugation time is preferably 3 to 300 minutes, more preferably 5 to 200 minutes, still more preferably 10 to 100 minutes, and still more preferably 15 to 60 minutes.
In addition, the centrifugation conditions are preferably 300 G or more, more preferably 500 G or more, and still more preferably 800 G or more, as the relative centrifugal acceleration, from the viewpoint of the recovery rate and purity of catechins and ease of separation. In addition, although an upper limit is not specifically limited, For example, 170000G or less is preferable, 120,000G or less is more preferable, and 70000G or less is still more preferable. The range of the relative centrifugal acceleration is preferably 300 to 170000G, more preferably 500 to 120,000G, and still more preferably 800 to 70000G. The rotation speed and the rotation radius of the centrifuge can be appropriately selected so that the relative centrifugal acceleration is within the above range.

The production method of the present invention can have the following characteristics (i) to (ii).
(I) In the purification method of the present invention, the yield of catechins is preferably 25% or more, more preferably 30% or more, and further preferably 35% or more.
(Ii) In the purified catechins-containing composition obtained by the production method of the present invention, the purity of catechins is preferably 25% by mass or more, more preferably 30% by mass or more, and further preferably 35% by mass or more. .

1. Analysis of catechins and caffeine A purified tea extract was appropriately diluted with a 0.1 mol / L acetic acid-dimethylsulfoxide solution and filtered through a 0.2 μm filter to prepare a sample. Catechins and caffeine are measured using a high performance liquid chromatograph (model SCL-10AVP, manufactured by Shimadzu Corporation), and a packed column for liquid chromatography using an octadecyl group (L-column TM ODS, 4.6 mmφ × 250 mm: Foundation Equipped with a chemical substance evaluation research organization), and a gradient method was performed at a column temperature of 35 ° C. As a standard product of catechins, a product manufactured by Mitsui Norin was used and quantified by a calibration curve method. The mobile phase A solution was a distilled aqueous solution containing 0.1 mol / L of acetic acid, the B solution was an acetonitrile solution containing 0.1 mol / L of acetic acid, the sample injection amount was 20 μL, and the UV detector wavelength was 280 nm. . The gradient conditions are as follows.

Time (minutes) Liquid A concentration (volume%) Liquid B concentration (volume%)
0.0 97 3
5.0 97 3
37.0 80 20
43.0 80 20
43.5 0 100
48.5 0 100
49.0 97 3
60.0 97 3

2. Recovery rate of catechins The recovery rate of catechins was calculated by the following formula (1).

    Recovery rate of catechins in purified catechins-containing composition = (mass of catechins contained in purified catechins-containing composition) / (mass of catechins contained in catechins-containing composition) × 100 (1)

3. Purity of catechins The purity of catechins was calculated by the following formula (2).
Purity of catechins of purified catechins-containing composition = (mass of catechins contained in purified catechins-containing composition) / (total solid content mass contained in purified catechins-containing composition) × 100 (2)

Preparation Example 1
0.12 g of green tea extract (catechin concentration 32% by mass) and 0.21 g of soybean phosphatidylcholine (Coatsome NC20, manufactured by NOF Corporation) as phospholipid were mixed with 10 g of ion-exchanged water. The mixture was incubated at 25 ° C. with shaking for 30 minutes. Subsequently, centrifugation was performed under the conditions of 25 ° C., 1000 G (rotation speed: 3000 rpm), and 30 minutes as a separation operation to precipitate the phospholipid phase and remove the supernatant liquid phase. The solid content of the precipitated phase was recovered to obtain a catechins-containing composition X.

Preparation Example 2
0.25 g of green tea extract (catechin concentration 32% by mass) and 0.81 g of soybean phosphatidylcholine (Coatsome NC20, manufactured by NOF Corporation) as phospholipid were mixed with 10 g of ion-exchanged water. The mixture was incubated at 25 ° C. with shaking for 30 minutes. Subsequently, centrifugation was performed under the conditions of 25 ° C., 1000 G (rotation speed: 3000 rpm), and 30 minutes as a separation operation to precipitate the phospholipid phase and remove the supernatant liquid phase. The solid content of the precipitated phase was recovered to obtain a catechin-containing composition Y.

Preparation Example 3
0.12 g of green tea extract (catechin concentration 32% by mass) and 0.20 g of soybean phosphatidylcholine (Coatsome NC20, manufactured by NOF Corporation) as phospholipid were mixed with 10 g of ion-exchanged water. The mixture was incubated at 25 ° C. with shaking for 30 minutes. Subsequently, centrifugation was performed under the conditions of 25 ° C., 1000 G (rotation speed: 3000 rpm), and 30 minutes as a separation operation to precipitate the phospholipid phase and remove the supernatant liquid phase. The solid content of the precipitated phase was recovered to obtain a catechins-containing composition Z.

Example 1
0.38 g of the catechins-containing composition X obtained in Preparation Example 1 and 10.02 g of a 20% by mass ethanol aqueous solution were mixed at 25 ° C. (the ethanol concentration after mixing was 19.7% by mass). The mixture was incubated at 25 ° C. with shaking for 30 minutes. Then, as a separation operation, centrifugation is performed under conditions of 25 ° C., 1000 G (rotation number 3000 rpm), 30 minutes, to precipitate a phase rich in phospholipids and contaminants, and recover a liquid phase rich in catechins in the supernatant, A purified catechin-containing composition was obtained. The obtained purified catechins-containing composition was analyzed. The results are shown in Table 1.

Examples 2-9
A purified catechin-containing composition was obtained in the same manner as in Example 1 except that an aqueous ethanol solution having the concentration shown in Table 1 was used. The obtained purified catechins-containing composition was analyzed. The results are shown in Table 1.

Comparative Example 1
A purified catechins-containing composition was obtained in the same manner as in Example 1 except that water was used instead of the ethanol aqueous solution. The obtained purified catechins-containing composition was analyzed. The results are shown in Table 1.

Example 10
A purified catechin-containing composition was obtained in the same manner as in Example 5 except that the amount of the aqueous ethanol solution shown in Table 2 was used. The obtained purified catechins-containing composition was analyzed. The results are shown in Table 2 together with the results of Example 5.

Examples 11-13
A purified catechin-containing composition was obtained in the same manner as in Example 5, except that the catechin-containing composition X was replaced with the catechin-containing composition Y, and an ethanol aqueous solution having an amount shown in Table 2 was used. The obtained purified catechins-containing composition was analyzed. The results are shown in Table 2 together with the results of Example 5.

Examples 14-17
Except that the catechin-containing composition X was replaced with the catechin-containing composition Z, the concentration of the ethanol aqueous solution was 40% by mass, and the catechin-containing composition and the ethanol aqueous solution were mixed at the temperatures shown in Table 3. In the same manner as in No. 10, a purified catechin-containing composition was obtained. The obtained purified catechins-containing composition was analyzed. The results are shown in Table 3.

  From Tables 1-3, from the catechins containing composition whose mass ratio [(B) / (A)] of (A) catechin and (B) phospholipid is 2-100, organic solvent aqueous solution, and organic solvent It can be seen that catechins can be recovered with high purity by mixing at least one selected solvent and recovering the catechin-rich phase generated in the mixed solution.

Claims (12)

(A) At least one selected from catechins-containing compositions having a mass ratio [(B) / (A)] of catechins to (B) phospholipids of 2 to 100, an organic solvent aqueous solution and an organic solvent Step A for mixing with the solvent of
Including a step B of recovering a phase rich in catechins generated in the mixed solution;
A method for producing a purified catechin-containing composition.   The manufacturing method of the refinement | purification catechin containing composition of Claim 1 whose content of (A) catechin in solid content of the said catechin containing composition is 1-35 mass%.   The purified catechin according to claim 1 or 2, wherein a mass ratio [(C) / (A)] of (A) catechins to (C) caffeine in the catechins-containing composition is 0.00001 to 1. Method for producing a kind-containing composition.   The manufacturing method of the refinement | purification catechin containing composition of any one of Claims 1-3 whose organic-solvent density | concentration in the said liquid mixture is 15-55 mass%.   The manufacturing method of the refinement | purification catechin containing composition of any one of Claims 1-4 whose organic-solvent density | concentration in the said organic-solvent aqueous solution is 10 mass% or more.   The manufacturing method of the refinement | purification catechin containing composition of any one of Claims 1-5 whose mass ratio of the total solvent amount with respect to the total amount of solids of the said catechin containing composition is 2-70.   The manufacturing method of the refined catechins containing composition of any one of Claims 1-6 whose said organic solvent is ethanol.   The manufacturing method of the refinement | purification catechin containing composition of any one of Claims 1-7 whose mixing temperature is 5-60 degreeC.   The method for producing a purified catechins-containing composition according to any one of claims 1 to 8, wherein the catechin-rich phase is a liquid phase produced by solid-liquid separation of the mixed solution.   The method for producing a purified catechins-containing composition according to claim 9, wherein the solid-liquid separation is centrifugation.   The method for producing a purified catechins-containing composition according to claim 10, wherein the centrifugation is performed under conditions of a temperature of 5 to 60 ° C, a relative centrifugal acceleration of 300 to 170,000 G, and a time of 3 to 300 minutes.   Before the said process A and the process B, a tea extract and a phospholipid are mixed, The process of collect | recovering the said catechins containing composition from the liquid mixture is included. Of producing a purified catechins-containing composition.