JP6730788B2 - Method for producing purified catechin-containing composition - Google Patents

Description

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

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

As a technique for reducing such contaminants, for example, tea leaves are extracted with hot water or an organic solvent aqueous solution, the solution containing the extracted components is washed with chloroform, and then the extracted components are redissolved in the organic solvent, and then the organic solvent is added. Method of distilling off (patent document 1), method of contacting tea extract with activated clay or acid clay (patent document 2), dissolving or suspending tea extract in water or hydrous organic solvent, and synthesizing under alkaline conditions A method of contacting with an adsorbent (Patent Document 3) and the like are known.

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

JP-A-59-219384 JP, 6-142405, A 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 in a higher yield.

The present inventor mixes a catechins-containing composition containing catechins and phospholipids in a specific ratio with a specific solvent to form a phase rich in catechins and a phase rich in phospholipids and impurities. It was found that catechins can be obtained in high purity and in a higher yield by recovering the separated and catechin-rich phase.

That is, the present invention provides a catechin-containing composition having a mass ratio [(B)/(A)) of (A) catechins and (B) phospholipids of 2 to 100, an aqueous organic solvent solution and an organic solvent. The present invention provides a method for producing a purified catechins-containing composition, which includes a step A of mixing at least one solvent selected from the group A and a step B of collecting a catechins-rich phase generated in the mixed solution. ..

According to the present invention, it is possible to provide a method for producing a purified catechins-containing composition capable of recovering catechins with high purity and in a higher yield.

The method for producing a purified catechin-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 in which the mass ratio [(B)/(A)] of (A) catechins and (B) phospholipids is 2 to 100, an organic solvent aqueous solution and an organic solvent. Is a step of mixing with at least one solvent.

Here, in the present specification, "catechins" means epigallocatechin gallate, gallocatechin gallate, a gallate body consisting of epicatechin gallate and catechin gallate, and a non-gallate body consisting of epigallocatechin, gallocatechin, epicatechin and catechin. Is also a generic term. The content of catechins is defined based on the total amount of the above eight kinds, and in the present invention, at least one of the above eight kinds of catechins may be contained.
Further, in the present specification, the term “phospholipid” is a general term for lipids having a phosphate ester moiety and a phosphonate ester moiety in the structure.

(Composition containing catechins)
The catechins-containing composition contains (A) catechins and (B) phospholipids and has a mass ratio [(B)/(A))] of 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 further preferable, and 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 the range of such a mass ratio, 2 to 100 is preferable, 4 to 50 is more preferable, 6 to 25 is further preferable, and 6 to 20 is even more preferable.

The content of (A) catechins in the solid content of the catechins-containing composition is preferably 1% by mass or more, more preferably 2% by mass or more, and 4% by mass or more from the viewpoint of the yield and the purity of the catechins. Is more preferable, and 35% by mass or less is preferable, 30% by mass or less is more preferable, and 25% by mass or less is further preferable. 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. Is. Here, in the present specification, the “solid content” is a residue obtained by drying the sample in an electric constant temperature dryer at 105° C. for 6 hours 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 of the catechin-containing composition is preferably 65% by mass or more, more preferably 70% by mass or more, and 75% by mass or more from the viewpoint of ease of separation operation. Is more preferable, and from the viewpoint of yield and purity of catechins, 99% by mass or less is preferable, 98% by mass or less is more preferable, and 96% by mass or less is further preferable. The range of the content of the (B) phospholipid is preferably 65 to 99% by mass, more preferably 70 to 98% by mass, and further preferably 75 to 96% by mass in the solid content of the catechin-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, Ltd.) or analysis using HPLC. ..

In the catechin-containing composition, the mass ratio [(C)/(A)] of (A) catechins and (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, but is preferably 0.00001 or more, more preferably 0.0001 or more. 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, still more preferably Is 0.0001 to 0.05.

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

Examples of the tea extract (A') include a tea extract, a concentrate thereof, and a purified product thereof, and various forms such as solids, liquids, solutions and slurries are available. The tea extract may be used alone or in combination of two or more. Here, in the present specification, the "tea extract" refers to one extracted from tea leaves using hot water or a hydrophilic organic solvent, which has not been concentrated or purified. As the extraction method and extraction conditions, known methods and conditions can be adopted, and there is no particular limitation. Examples of the tea leaves include Camellia genus, for example, C. sinensis var. sinensis (including Yabukita 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 depending on the processing method.
Examples of the non-fermented tea include green tea such as sencha, bancha, tencha, tea in a pot, stem tea, stick tea, and bud tea. In addition, examples of the semi-fermented tea include oolong tea such as iron Kannon, color varieties, golden katsura, and Wuyiwa rock tea. Furthermore, examples of fermented tea include black tea such as Darjeeling, Assam, and Sri Lanka. These can be used alone or in combination of two or more. Among them, non-fermented tea is preferable, and green tea is more preferable, from the viewpoint of the content of catechins.

Further, the "concentrate of tea extract" is one in which a part of the solvent is removed from the tea extract to increase the concentration of catechins. For example, the concentration method includes atmospheric concentration, reduced pressure concentration, and membrane concentration. Etc. can be mentioned. A commercially available product may be used as the concentrate of the tea extract, for example, green tea such as "Polyphenon" from Mitsui Norin Co., Ltd., "Theafran" from ITO EN Co., Ltd., "Sunphenon" from Taiyo Kagaku Co., Ltd. An extract concentrate may be mentioned. Further, the "purified product of tea extract" is a product obtained by purifying a tea extract or its concentrate to increase the purity of catechins, for example, JP-A-2004-147508 and JP-A-2004-149416. The methods described in Japanese Patent Laid-Open No. 2006-160656, Japanese Patent Laid-Open No. 2007-252568, Japanese Patent Laid-Open No. 2008-079609 and the like can be adopted. In addition, the tea extract may be subjected to various chromatographies such as reverse phase chromatography and liquid chromatography to fractionate catechins.

The tea extract is preferably in the form of an aqueous solution. The aqueous solution in which the tea extract is dissolved is, for example, a tea extract extracted from tea leaves with water, diluted with water or concentrated as necessary, and then used to concentrate the tea extract or a purified product thereof with water. The tea extract, its concentrate, or the dried product of the purified product thereof may be diluted with water and used again.

The content of catechins in the aqueous solution in which the tea extract is dissolved can be appropriately selected, but from the viewpoint of purification efficiency, 0.02 mass% or more is preferable, 0.05 mass% or more is more preferable, and 0. 06 mass% or more is more preferable, 10 mass% or less is preferable, 7 mass% or less is more preferable, and 5 mass% or less is further preferable. The range of the content of such catechins is preferably 0.02 to 10% by mass, more preferably 0.05 to 7% by mass, and further preferably 0.06 to 5% by mass.

In the tea extract used in the present invention, the proportion of gallate bodies in catechins (hereinafter, also referred to as “gallate body proportion”) is 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 further preferable. The range of the percentage of gallate is preferably 30 to 70% by mass, more preferably 35 to 65% by mass, and further preferably 40 to 60% by mass. Here, in the present specification, the “gallate body ratio” is a mass ratio of the above-mentioned four gallate bodies to eight catechins.

(B) As the 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 And so on. The constituent fatty acid of the phospholipid (B) is not particularly limited and may be either saturated fatty acid or unsaturated fatty acid. In addition to the neutral phospholipid, the (B) phospholipid may include a charged phospholipid such as an anionic phospholipid and a cationic phospholipid, and further a polymerizable phospholipid. The phospholipids can be used alone or in combination of two or more.

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

The phospholipid when mixed with the tea extract may be in the form of a phospholipid membrane or a liposome. The phospholipid in the form of a phospholipid membrane or a liposome facilitates the incorporation of catechins into the phospholipid membrane or its internal aqueous phase and the adsorption of catechins to the phospholipid membrane. As used herein, the term "liposome" refers to a closed vesicle having an inner aqueous phase portion surrounded by a phospholipid bilayer membrane, and the multiphase liposome (Multilamellar Vesicle: MLV), large unilamellar vesicles (LUV), and small unilamellar vesicles (SUV). Any type of liposome can be used in the present invention.

When adsorbing catechins to a phospholipid membrane, for example, a tea extract and phospholipid are mixed, and stress is applied by shaking or the like to form a phospholipid membrane and adsorption of non-polymer catechins to the phospholipid membrane. And may be performed substantially at the same time. It is also possible to apply stress to the phospholipid by shaking or the like to form a phospholipid film in advance, and then mix the tea extract with the phospholipid film.
The method for forming liposomes is not particularly limited, and methods such as Bangham method, lipid dissolution method, mechanochemical method, and freeze-dried liposome method can be adopted.

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, still more preferably from the viewpoint of the separation operation between the phospholipid phase and the liquid phase. The thickness is 50 to 5,000 nm, more preferably 50 to 3,000 nm, even more preferably 80 to 1,000 nm, and particularly preferably 100 to 500 nm. The term "median diameter" as used herein means a particle diameter corresponding to a cumulative value of 50% (d ₅₀ ) in a volume-based cumulative particle size distribution measured by a laser diffraction scattering method.

In addition, the phospholipid when mixed with the tea extract preferably has a phase transition temperature. Here, the “phase transition temperature” in the present specification refers to a temperature at which a phase transition between the gel and the liquid crystal that the phospholipid can take occurs, and when a sufficient amount of water is present with respect to the phospholipid. Is the value of. The phase transition temperature can be measured by a differential thermal analysis using a differential scanning calorimeter (DSC).
As the differential scanning calorimeter, for example, DSC7020 (manufactured by Hitachi High-Tech Science) can be used. Specifically, the phase transition temperature of phospholipid can be measured by the following procedure.
(1) For phospholipids having a phase transition temperature above 0° C., the phase transition temperature is measured by the following procedure. First, a sample is prepared in which phospholipids are sufficiently dispersed in water so that a sufficient amount of water is present with respect to the phospholipids and the phospholipid concentration is 1 to 10 g/100 g. Next, 0.005 g of the sample is weighed in an aluminum sample container, covered with an aluminum cover, and then sealed using an electric sample sealer. Place the sealed sample container on the holder unit in the electric furnace. Further, the blank container in which air is sealed is placed on the blank side of the holder unit. Cover the electric furnace and hold under a nitrogen atmosphere at 1° C. for 5 minutes. Then, heating is performed from 1° C. to 98° C. at a heating rate of 0.5° C./min (heating step). In this heating 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 the endothermic peak is observed in the temperature raising step is the phase transition temperature of the phospholipid.
(2) For phospholipids whose phase transition temperature cannot be confirmed under the above measurement conditions, the phase transition temperature is measured by the following procedure. First, a 50% by mass ethylene glycol aqueous solution with respect to phospholipid is prepared as a sample in which the phospholipid is sufficiently dispersed in water so that the phospholipid concentration becomes 1 to 10 g/100 g. Then, a sample container and a blank container are prepared by the same operation as above, and placed on the holder unit. Cover the electric furnace and hold at 25° C. for 5 minutes under a nitrogen atmosphere. Then, it is cooled from 25° C. to −40° C. at a temperature lowering rate of 0.5° C./min (cooling step). 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 the exothermic peak is observed in the cooling step is the phase transition temperature of the phospholipid.

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

Examples of the phospholipid having a phase transition temperature include soybean phosphatidylcholine (phase transition temperature measured under the measurement conditions in the present specification of about −10° C.), hydrogenated soybean phosphatidylcholine (phase transition temperature measured under the measurement conditions in the present specification). About 51° C.), dimyristoylphosphatidylcholine (phase transition temperature about 23° C. described in the following reference 1), dipalmitoylphosphatidylcholine (phase transition temperature about 41° C. described in the following reference 1), distearoylphosphatidylcholine (reference below) Reference 1 describes a phase transition temperature of about 54° C., dihexadecylphosphatidylcholine (reference reference 2 below describes a phase transition temperature about 45° C.), stearoylpalitomylphosphatidylcholine (reference 3 below describes a phase transition temperature about 45). C.), dipalmitoylphosphatidylethanolamine (phase transition temperature of about 63.degree. C. described in Reference 2 below), 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, but a phospholipid content of 60% by mass or more is preferable, and a phospholipid content of 80% by mass or more is preferable. More preferable.

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

From the viewpoint of production efficiency, the mixing ratio of the tea extract and the phospholipid is 0 as the mass ratio [(A)/(B)] of the (A) catechins and the (B) phospholipid in the tea extract. 0.001 or more is preferable, 0.01 or more is more preferable, 0.02 or more is further preferable, and from the viewpoint of yield and purity of catechins, 0.20 or less is preferable, 0.18 or less is more preferable, 0 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. It is preferably 0.02 to 0.09.

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

The pH (25° C.) at the time of mixing the tea extract and the phospholipid is preferably 2.5 to 9.0, more preferably 4.0 to 8 from the viewpoint of equipment corrosion prevention and stability of catechins. 0.0, and 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 added at the same time and mixed, or one may be added to the other and mixed. Further, when mixing the tea extract and the phospholipid, treatments such as stirring, shaking, and ultrasonic irradiation may be performed.

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 contaminants 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 mixed solution, and the recovery method includes solid-liquid separation such as ultrafiltration, centrifugation, gel chromatography and dialysis. Solid-liquid separation can be performed by one kind or a combination of two or more kinds. Of these, centrifugation is preferred. By centrifuging the mixed solution, the phospholipid phase can be easily recovered as a precipitation phase.

As the centrifuge, a general equipment such as a separation plate type, a cylinder type, a decanter type or the like can be used, and an ultracentrifuge may be used if 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, still more preferably 15 to 60 minutes, from the viewpoint of the recovery rate of the phospholipid phase.
In addition, the conditions of centrifugation are, for example, 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 the precipitation phase. Is more preferable. The upper limit is not particularly limited, but is preferably 170,000 G or less. The range of the relative centrifugal acceleration is preferably 300 to 170000 G, more preferably 500 to 170000 G, and further preferably 800 to 170000 G. The rotation speed and the rotation radius of the centrifuge can be appropriately selected so that the relative centrifugal acceleration falls within the above range. Here, in the present specification, the “relative centrifugal acceleration” means 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 thus recovered can be used as it is as the catechin-containing composition in this 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 thereof include alcohols such as ethanol and methanol, ketones such as acetone, and esters such as ethyl acetate. Of these, alcohols and ketones are preferable, and alcohols are more preferable and ethanol is still more preferable in consideration of use in foods.

From the viewpoint of the yield and purity of catechins, the concentration of the organic solvent in the aqueous organic solvent solution is preferably 10% by mass or more, more preferably 20% by mass or more, further preferably 30% by mass or more, and more preferably 40% by mass or more. More preferable.

(mixture)
The method for mixing the catechin-containing composition and the solvent is not particularly limited, and the catechin-containing composition and the solvent may be added at the same time and mixed, or one may be added to the other and mixed. Further, when the catechin-containing composition and the solvent are mixed, treatments such as stirring, shaking, and ultrasonic irradiation may be performed.

From the viewpoint of yield and purity of catechins, the amount of the solvent used is preferably 2 or more, more preferably 4 or more, and more preferably 6 or more as a mass ratio of the total amount of the solvent to the total amount of solids in the catechins-containing composition. From the viewpoint of productivity, 70 or less is preferable, 60 or less is more preferable, and 50 or less is further preferable. The mass ratio range is preferably 2 to 70, more preferably 4 to 60, and further preferably 6 to 50. The term "total amount of solvent" as used herein means the sum of the amount of solvent (for example, water) contained in the catechin-containing composition and the amount of organic solvent aqueous solution or organic solvent used for mixing.

From the viewpoint of the yield of catechins, the organic solvent concentration in the mixed liquid 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 further preferable. The range of such organic solvent concentration is preferably 15 to 55% by mass, more preferably 20 to 53% by mass, and further 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 a catechin-containing composition and an organic solvent aqueous solution, and adjusting the organic solvent concentration within the above range, the catechin-containing composition is water. After dissolving in, the method of adjusting the organic solvent concentration within the above range by adding an organic solvent, after dispersing or dissolving the catechin-containing composition in the organic solvent, gradually add water to the organic solvent concentration within the above range The method of adjusting to. 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. May be 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, further preferably 20° C. or higher, particularly preferably 30° C. or higher, from the viewpoint of the recovery rate and purity of the catechins. From the viewpoint of the stability of catechins, 60°C or lower is preferable, 55°C or lower is more preferable, and 50°C or lower is further preferable. The range of the mixing temperature is preferably 5 to 60°C, more preferably 10 to 55°C, further preferably 20 to 55°C, particularly preferably 30 to 50°C.
The mixing time of the catechins-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 the catechins, the purification efficiency, 10 minutes or more is preferable, and 15 minutes or more is more preferable. 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 further preferable. The range of such 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 mixed liquid is kept constant for a predetermined time.

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

As the centrifuge, a general equipment such as a separation plate type, a cylinder type, a decanter type or the like can be used, and an ultracentrifuge may be used if 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 preferably 30° C. or higher, from the viewpoint of the yield of catechins and the ease of separation. From the viewpoint of the stability of catechins, 60°C or lower is preferable, 55°C or lower is more preferable, and 50°C or lower is further preferable. The temperature range is preferably 5 to 60° C., more preferably 10 to 55° C., further preferably 20 to 55° C., and particularly preferably 30 to 50° C.
The centrifugation time is preferably 3 minutes or more, more preferably 5 minutes or more, still more preferably 10 minutes or more, still more preferably 15 minutes or more, from the viewpoints of the recovery rate and purity of catechins and the ease of separation. And 300 minutes or less is preferable, 200 minutes or less is more preferable, 100 minutes or less is further preferable, and 60 minutes or less is further more preferable. The range of centrifugation time is preferably 3 to 300 minutes, more preferably 5 to 200 minutes, still more preferably 10 to 100 minutes, still more preferably 15 to 60 minutes.
The centrifugation conditions are, for example, 300 G or more, preferably 500 G or more, more preferably 800 G or more, as the relative centrifugal acceleration, from the viewpoints of the recovery rate and purity of catechins and the ease of separation. Although the upper limit is not particularly limited, for example, 170,000 G or less is preferable, 120000 G or less is more preferable, and 70,000 G or less is further preferable. The range of the relative centrifugal acceleration is preferably 300 to 170000 G, more preferably 500 to 120,000 G, and further preferably 800 to 70,000 G. The rotation speed and the rotation radius of the centrifuge can be appropriately selected so that the relative centrifugal acceleration falls within the above range.

The manufacturing 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, still more preferably 35% or more.
(Ii) The purified catechin-containing composition obtained by the production method of the present invention has a catechin purity of 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 The purified tea extract was appropriately diluted with a 0.1 mol/L acetic acid-dimethyl sulfoxide solution and filtered with a 0.2 μm filter to prepare a sample. The catechins and caffeine were measured using a high performance liquid chromatograph (model SCL-10AVP, manufactured by Shimadzu Corporation), and a packed column for an octadecyl group-introduced liquid chromatograph (L-column TM ODS, 4.6 mmφ×250 mm: Foundation). A corporation chemical substance evaluation research institute) was attached, and the column temperature was 35° C. and the measurement was carried out by a gradient method. As the standard catechins, those manufactured by Mitsui Norin were used and quantified by the calibration curve method. The mobile phase solution A was a distilled aqueous solution containing 0.1 mol/L of acetic acid, the solution B 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
43.50 100
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 the purified catechins-containing composition=(mass of catechins contained in the purified catechins-containing composition)/(mass of catechins contained in the catechins-containing composition)×100 (1)

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

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

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

Preparation Example 3
0.12 g of green tea extract (catechins concentration 32% by mass) and 0.20 g of soybean phosphatidylcholine (Coatsome NC20, manufactured by NOF CORPORATION) as phospholipids were mixed with 10 g of ion-exchanged water. The mixture was incubated at 25°C with shaking for 30 minutes. Next, as a separating operation, centrifugation was performed under the conditions of 25° C., 1000 G (rotation speed 3000 rpm), and 30 minutes to precipitate the phospholipid phase and remove the supernatant liquid phase. The solid content of the precipitation phase was recovered to obtain a catechin-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 mass% ethanol aqueous solution were mixed at 25° C. (the ethanol concentration after mixing was 19.7 mass %). The mixture was incubated at 25°C with shaking for 30 minutes. Then, as a separation operation, centrifugation is performed under the conditions of 25° C., 1000 G (rotation speed 3000 rpm), and 30 minutes to precipitate a phase rich in phospholipids and contaminants, and a supernatant catechin-rich liquid phase is recovered. 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 the aqueous ethanol solution having the concentrations 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 catechin-containing composition was obtained by the same operation as in Example 1 except that water was used instead of the aqueous ethanol 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 catechins-containing composition was obtained in the same manner as in Example 5, except that the catechins-containing composition X was replaced with the catechins-containing composition Y and 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 14-17
Examples except that the catechins-containing composition X was replaced with the catechins-containing composition Z, the concentration of the ethanol aqueous solution was 40% by mass, and the catechins-containing composition and the ethanol aqueous solution were mixed at the temperature shown in Table 3 By the same operation as in 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 to 3, 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, It can be seen that the catechins can be recovered in high purity by mixing with at least one selected solvent and recovering the catechins-rich phase generated in the mixed solution.

Claims (11)

A catechin-containing composition having a mass ratio [(B)/(A)] of (A) catechins and (B) phospholipids of 2 to 100, and at least one solvent selected from an aqueous alcohol solution and alcohol And a step A in which the mass ratio of the total solvent amount to the total solid content is 2 to 70 ,
Including a step B of recovering a catechin-rich phase generated in the mixed solution,
A method for producing a purified catechin-containing composition. The method for producing a purified catechins-containing composition according to claim 1, wherein the content of (A) catechins in the solid content of the catechins-containing composition is 1 to 35% by mass. The purified catechin according to claim 1 or 2, wherein a mass ratio [(C)/(A)] of (A) catechins and (C) caffeine in the catechin-containing composition is 0.00001 to 1. Of producing a composition containing compounds. The method for producing a purified catechin-containing composition according to any one of claims 1 to 3, wherein the alcohol concentration in the mixed solution is 15 to 55% by mass. The method for producing a purified catechin-containing composition according to any one of claims 1 to 4, wherein the alcohol concentration in the alcohol aqueous solution is 10% by mass or more. Wherein the alcohol is ethanol, the production method of the purified catechins-containing composition according to any one of claims 1-5. Mixing temperature is 5 to 60 ° C., the manufacturing method of the purified catechins-containing composition according to any one of claims 1-6. Manufacturing method of the phase rich in catechins, wherein a resulting liquid phase by mixing liquid to solid-liquid separation, any one of purified catechins-containing composition according to claim 1-7. The method for producing a purified catechin-containing composition according to claim 8 , wherein the solid-liquid separation is centrifugation. The method for producing a purified catechin-containing composition according to claim 9 , wherein the centrifugation is performed at a temperature of 5 to 60° C., a relative centrifugal acceleration of 300 to 17,000 G, and a time of 3 to 300 minutes. Prior to the step A and step B, and mixing the tea extract and phospholipid, a mixed solution comprising the step of recovering the catechins-containing composition, according to any one of claims 1-10 A method for producing a purified catechin-containing composition according to claim 1.