JP4866815B2 - Method for producing purified green tea extract - Google Patents

Description

  The present invention relates to a method for producing a purified green tea extract.

  As a physiological effect of catechin, an α-amylase activity inhibitory action and the like have been reported (for example, see Patent Document 1). In order to express such a physiological effect, since it is necessary to ingest a large amount of catechin more simply, a method of blending catechin with a high concentration in a beverage has been desired.

  As one of the methods, a method of adding catechin to a beverage in a dissolved state using a tea extract such as a concentrate of green tea extract (Patent Document 2) is used. However, depending on the type of beverage for which catechin is added at a high concentration by this method, for example, when catechin is added to black tea extract or carbonated beverage, the remaining bitterness derived from caffeine and green tea is a beverage product. It has been found to greatly detract from value.

As a method for removing impurities such as caffeine from the green tea extract, an adsorption method (Patent Documents 2 to 7), an extraction method (Patent Documents 8 and 9), and the like are known.
In the above method, when increasing the content of non-polymer catechins in the green tea extract, it is necessary to use an organic solvent. However, when viewed industrially, there is a problem that the recovery rate is low. . Moreover, when using alkaline aqueous solution, when mix | blending with a drink, there existed a subject that the water-insoluble component derived from a tea leaf remained, and there was no effective means with respect to this.

As a method for improving the color tone of a green tea extract, an antibacterial deodorant (Patent Document 10) is known which is obtained by allowing activated carbon to act on a tea extract in the presence of cyclodextrin and adsorbing and removing colored components on the activated carbon. However, its use in beverage catechin preparations has been difficult.
JP-A-3-133828 JP-A-5-153910 JP-A-8-109178 JP 2002-335911 A JP 2006-36645 A JP-A-1-175978 JP 2001-97968 A JP-A-1-289447 JP 59-219384 A JP 2001-299887 A

In addition to the conventionally known impurities such as caffeine, bitterness, and sourness reduction, the present inventors have found that the reduction of the thickening and nitrogen content has the effect of improving the taste of the purified green tea extract. I found it.
The object of the present invention is to extract a purified green tea containing a high concentration of non-polymer catechins, in which impurities such as caffeine are removed, bitterness, acidity, thickness, nitrogen content is reduced, and hue is good It is in providing the manufacturing method of a thing.

  As a result of studying the purification treatment of non-polymer catechins in the green tea extract, the present inventors passed the green tea extract through the synthetic adsorbent and once adsorbed to the synthetic adsorbent. Contact with an aqueous solution, wash the synthetic adsorbent, and then contact with an organic solvent aqueous solution to recover the eluted non-polymer catechins, thereby reducing caffeine and bitterness, sourness, thickening, It has been found that a purified green tea extract having a high concentration of non-polymer catechins with reduced nitrogen content and hue can be obtained.

  That is, the present invention adsorbs a green tea extract to a synthetic adsorbent, contacts the synthetic adsorbent with a basic aqueous solution, cleans the synthetic adsorbent, contacts an organic solvent aqueous solution, and elutes the non-polymer. The present invention provides a method for producing a purified green tea extract that recovers catechins.

  According to the present invention, a purified green tea extract containing a high concentration of non-polymer catechins in which impurities such as caffeine are removed, bitterness, sourness, thickness and nitrogen content are reduced and the hue is good Is advantageously obtained industrially.

  In the present invention, the non-polymer catechins include non-epide catechins such as catechin, gallocatechin, catechin gallate and gallocatechin gallate and epi-catechins such as epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate. It is a general term.

  In the present invention, the non-polymer catechin gallate body is a general term including catechin gallate, gallocatechin gallate, epicatechin gallate, epigallocatechin gallate and the like.

  Examples of the green tea extract used in the present invention include an extract obtained from green tea leaves. More specifically, the tea leaves used include tea leaves made from tea leaves obtained from the genus Camellia, for example, C. sinensis, C. assamica, and camellia seeds or hybrids thereof. The tea leaves produced include green teas such as sencha, bancha, gyokuro, tencha, and kettle roasted tea. Moreover, you may use the tea leaf which gave the carbon dioxide contact process of the supercritical state.

  Extraction from green tea leaves is performed by stirring extraction or the like using water or a water-soluble organic solvent or a mixture thereof as an extraction solvent. In the extraction, an organic acid salt such as sodium ascorbate or an organic acid may be added in advance to water or a water-soluble organic solvent or a mixture thereof. Moreover, you may use together the method of extracting in so-called non-oxidative atmosphere, ventilating inert gas, such as boiling deaeration and nitrogen gas, and removing dissolved oxygen. The extract thus obtained can be used in the present invention as it is, even if it is dried and concentrated. Examples of the green tea extract include liquid, slurry, semi-solid, and solid state.

For the green tea extract used in the present invention, instead of using the extract extracted from the green tea leaves, the concentrate of the green tea extract may be dissolved in water or an organic solvent or diluted in water or an organic solvent, You may use together the extract from a green tea leaf, and the concentrate of a green tea extract.
Here, the concentrate of the green tea extract is obtained by concentrating an extract extracted from green tea leaves with hot water or a water-soluble organic solvent. For example, JP-A-59-219384, JP-A-4- This refers to those prepared by the methods described in JP-A-20589, JP-A-5-260907, JP-A-5-306279, and the like. Specifically, commercially available crude catechin preparations such as “Polyphenone” manufactured by Tokyo Food Techno Co., “Theafuran” manufactured by ITO EN, “Sunphenon” manufactured by Taiyo Kagaku Co., Ltd. can be used as the solid green tea extract.

  Moreover, it is preferable that the green tea extract is hydrolyzed before being adsorbed to the synthetic adsorbent from the viewpoint of reducing the gallate content in non-polymer catechins and reducing bitterness. The concentration reduction of the non-polymer catechin gallate body in the non-polymer catechins by hydrolysis is preferably 5% by mass or more, more preferably 7% by mass or more, and particularly preferably 10% by mass or more from the viewpoint of improving taste. The hydrolysis method is carried out by treatment with enzymes, acid treatment, alkali treatment or the like. As the enzyme, an enzyme having tannase activity, a bacterial cell or a culture solution, hydrochloric acid, sulfuric acid, phosphoric acid as the acid, and caustic soda as the alkali are preferable. Among them, hydrolysis with enzymes is preferable from the viewpoint of reaction control. Here, having tannase activity means having activity of degrading tannin, and any enzyme, fungus body, or culture solution can be used as long as it has this activity.

Specifically, pectinase PL Amano (manufactured by Amano Enzyme), hemicellulase amano 90 (manufactured by Amano Enzyme), tannase KTFH (manufactured by Kikkoman), etc. can be used as commercially available enzymes having tannase activity. Of these, tannase is preferred. For example, tannase obtained by culturing tannase-producing bacteria belonging to the genus Aspergillus, Penicillium or Rhizopus. Of these, those derived from Aspergillus oryzae are preferred.
The microbial cell having tannase activity is a microbial cell capable of producing an enzyme having tannase activity, and examples thereof include koji molds. Specific examples include the genus Aspergillus and the genus Penicillium. Of these, Aspergillus oryzae is preferred.
The culture solution having tannase activity is a culture solution obtained by culturing a tannase-producing bacterium of the genus Aspergillus, Penicillium, or Rhizopus. Preferably, a culture solution obtained by culturing tannic acid as the sole carbon source can be mentioned, and it can be used regardless of whether it is a purified product or an unpurified product.
Hydrolysis is preferably completed in as short a time as possible from the viewpoint of suppression of thickening deterioration and productivity, and it is preferable to use an enzyme or a culture solution.
The enzyme or culture solution having tannase activity used in the present invention preferably has an enzyme activity of 500 to 100,000 U / g, and is treated within a limited time industrially as 500 U / g or more. The enzyme reaction rate can be controlled to be 100,000 U / g or less. Here, 1 Unit represents the amount of enzyme that hydrolyzes 1 micromole of an ester bond contained in tannic acid in water at 30 ° C.

The non-polymer catechin concentration in the treatment with an enzyme having tannase activity and a culture solution is preferably 0.1 to 22% by mass, more preferably 0.1 to 15% by mass, and particularly preferably 0.5 to 10%. % By weight, particularly preferably 0.5 to 3% by weight. If it is 0.1% by mass or more, the amount of adsorption increases at the time of subsequent adsorption to the synthetic adsorbent, and if it is 22% by mass or less, the hydrolysis treatment is shortened, and the productivity and the taste of the green tea extract are obtained. To preferred.
In order to obtain a non-polymer catechin gallate body ratio with improved taste, an enzyme or a culture solution is added to the non-polymer catechins in the green tea extract so as to be in the range of 0.01 to 10% by mass. Is preferred. In order to complete the hydrolysis treatment within 2 hours, which is an industrially optimal enzyme reaction time, including the enzyme deactivation step, the enzyme or culture solution concentration is 0.01 to 7% by mass, and further, It is preferable that it is 03-5 mass%.
An enzyme or culture solution having tannase activity for non-polymer catechins in green tea extract is preferably 1 to 300 Unit / g-non-polymer catechin, more preferably 3-200 Unit / g-non-polymer catechin, Especially preferably, it adds so that it may become 5-150Unit / g-non-polymer catechin.
The treatment temperature with the enzyme or the culture solution is preferably 0 to 70 ° C., more preferably 0 to 60 ° C., and particularly preferably 5 to 50 ° C. at which the optimum enzyme activity can be obtained.

In order to terminate the hydrolysis reaction with the enzyme or the culture solution, it is necessary to deactivate the enzyme. Enzyme inactivation is achieved by heating. The enzyme deactivation temperature is preferably 70 to 100 ° C. If it is less than 70 ° C., the enzyme cannot be inactivated sufficiently in a short time, so that the hydrolysis reaction proceeds.
The enzyme deactivation method can be stopped by heating in a batch system or a continuous system such as a plate heat exchanger. In addition, after completion of inactivation of tannase, the tea extract can be clarified by an operation such as centrifugation.

  When, for example, gonococcus is used as the bacterial cell, the concentration of non-polymer catechins is preferably 0.1 to 22% by mass, more preferably 0.1 to 15% by mass, and particularly preferably 0.5 to 15% by mass. % Is added to the tea extract and hydrolyzed. Aspergillus is in the range of 0.5% by mass to 10% by mass with respect to the non-polymer catechins in the tea extract depending on the type and the like. Particularly preferably, it is added in the range of 1.0% by mass to 5% by mass. As temperature conditions, 45 to 70 degreeC, Furthermore, 50 to 60 degreeC is preferable. The fermentation time is preferably 12 hours to 20 days, and more preferably 1 day to 10 days. The inactivation of the enzyme activity of Aspergillus is the same as when the hydrolysis reaction with the enzyme or the culture solution is terminated.

In the present invention, the green tea extract is first adsorbed on the synthetic adsorbent. Caffeine and gallic acid can be reduced by the synthetic adsorbent treatment.
After adsorption, it is preferable to wash the synthetic adsorbent and remove gallic acid and impurities in the synthetic adsorbent before contacting the basic aqueous solution.
Synthetic adsorbents are generally insoluble three-dimensional crosslinked structure polymers that are substantially free of functional groups such as ion exchange groups. Preferably, those having an ion exchange group of less than 1 meq / g can be used. As the synthetic adsorbent used in the present invention, the matrix is styrene, such as Amberlite XAD4, XAD16HP, XAD1180, XAD2000 (Rohm & Haas, USA); Diaion HP20, HP21 (Mitsubishi Chemical); Sepabead SP850, SP825, SP700, SP70 (manufactured by Mitsubishi Chemical Corporation); VPOC1062 (manufactured by Bayer Corporation), modified styrene series in which the bromine atom is replaced with a nucleus to enhance the adsorptive power, such as Sepabeads SP205, SP206, SP207 (manufactured by Mitsubishi Chemical Corporation), methacrylic Systems such as Diaion HP1MG, HP2MG (Mitsubishi Chemical), phenols such as Amberlite XAD761 (Rohm and Haas), acrylics such as Amberlite XAD7HP (Rohm and Haas), polyvinyl, If example TOYOPEARL, HW-40C (manufactured by Tosoh Corporation), dextran-based, for example SEPHADEX, LH-20 (manufactured by Pharmacia), and the like can be used.
As the synthetic adsorbent, the matrix is preferably styrene, methacrylic, acrylic, or polyvinyl, and styrene is particularly preferable from the viewpoint of separability between catechin and caffeine.

As a means for adsorbing green tea extract to synthetic adsorbent, use a batch method in which synthetic adsorbent is added to green tea extract, stirred and adsorbed, and then recovered by filtration, or a column packed with synthetic adsorbent is used. A column method in which adsorption treatment is performed by continuous treatment is employed, but a continuous treatment method using a column is preferred from the viewpoint of productivity.
The column packed with the synthetic adsorbent is preferably washed in advance with water or an organic solvent aqueous solution. Examples of the water-soluble organic solvent include acetone, methanol, ethanol and the like, and ethanol is preferable from the viewpoint of use in foods. The concentration of the organic solvent to be contained is preferably 0 to 20% by mass, preferably 0 to 10% by mass, and more preferably 0 to 5% by mass from the viewpoint of catechin recovery. Preliminarily washed with 95 vol% aqueous ethanol solution under conditions of SV (space velocity) = 0.5 to 10 [h ⁻¹ ] and 2 to 10 [v / v] as the passing ratio for the synthetic adsorbent, and then synthetically adsorbed It is preferable to remove the raw material monomer of the agent and impurities in the raw material monomer. Then, SV = 0.5 to 10 [h ⁻¹ ], and a water passage condition of 1 to 60 [v / v] as a liquid passage ratio with respect to the synthetic adsorbent is performed to remove ethanol and remove the synthetic adsorbent. The ability to adsorb non-polymer catechins is improved by replacing the liquid-containing solution with an aqueous system.

As a means for adsorbing the green tea extract onto the synthetic adsorbent, it is preferable to pass the tea extract through a column packed with the synthetic adsorbent. The conditions for passing the green tea extract through the column filled with the synthetic adsorbent were SV (space velocity) = 0.5 to 10 [h ⁻¹ ], and 0 as the passage ratio for the synthetic adsorbent. It is preferable to pass through at 5 to 20 [v / v]. Adsorption of non-polymer catechins may be insufficient or unstable when the flow rate is 10 [h ^-1 ] or more and the flow rate is 20 [v / v] or more.

After adsorbing the green tea extract to the synthetic adsorbent, a basic aqueous solution is contacted. Thereby, the gallic acid and impurities adhering to the synthetic adsorbent can be removed. In some cases, the entire amount of the cleaning solution may be discarded from the viewpoint of removing impurities, or may be partially recovered from the viewpoint of improving the yield. As the basic aqueous solution to be used, alkali metal salts, alkaline earth alkaline aqueous solutions, preferably sodium-based alkaline aqueous solutions such as sodium hydroxide aqueous solution and sodium carbonate aqueous solution can be suitably used. The pH of the alkaline aqueous solution is preferably in the range of 7-11. From the viewpoint of non-polymer catechins recovery and removal of thickening components, 8 to 10.5, particularly 8.5 to 10 is preferable. Examples of the sodium-based aqueous solution having a pH of 7 to 11 include a 4% or less sodium hydroxide aqueous solution and a 4% or less sodium carbonate aqueous solution. The basic aqueous solution may contain a water-soluble organic solvent. A water-soluble organic solvent can be used as the organic solvent. Examples of the water-soluble organic solvent include acetone, methanol, ethanol and the like, and ethanol is preferable from the viewpoint of use in foods. The concentration of the organic solvent to be contained is preferably 0 to 20% by mass, preferably 0.1 to 10% by mass, and more preferably 1 to 5% by mass from the viewpoint of catechin recovery.
Synthetic adsorbent with a flow rate of SV (space velocity) = 0.5-10 [h ⁻¹ ] and a flow rate of 0.5-10 [v / v] with respect to the synthetic adsorbent. It is preferable to remove gallic acid and impurities adhering to the surface. Furthermore, contact at a flow rate of SV = 1.0 to 5 [h ^-1 ] and a flow rate of 1 to 5 [v / v] can remove gallic acid and impurities and recover non-polymer catechins. It is preferable in terms of rate.

  An organic solvent aqueous solution is further contacted with the synthetic adsorbent after contact with the basic aqueous solution to elute non-polymer catechins from the synthetic adsorbent, and the eluate is recovered. Examples of the organic solvent used here include acetone, methanol, ethanol and the like, and ethanol is preferable from the viewpoint of use in foods. The concentration of the organic solvent aqueous solution is 5 to 50%, further 10 to 45%, and especially 15 to 40%. The recovery rate of non-polymer catechins, bitterness, sourness, thickness, nitrogen content and hue reduction effect From the point of view, it is preferable.

The contact treatment is preferably carried out by passing through the column. As the flow-through conditions, the flow rate of SV (space velocity) = 0.5 to 10 [h ⁻¹ ] and 0 as the flow-through multiple for the synthetic adsorbent. It is preferable to remove gallic acid and impurities adhering to the synthetic adsorbent at a rate of 0.5 to 10 [v / v]. Furthermore, it is possible to remove gallic acid and impurities and recover non-polymer catechins by washing at a flow rate of SV = 1.0 to 5 [h ⁻¹ ] and a flow rate of 1 to 5 [v / v]. It is preferable in terms of rate.

  Furthermore, as a way of contact with the significant solvent aqueous solution, it may be eluted by bringing the organic solvent aqueous solution (2) into contact with the organic solvent aqueous solution (1). When elution is performed using such a concentration gradient, the concentration of the organic solvent aqueous solution (1) is 5 to 50%, more preferably 10 to 30%, and the concentration of the organic solvent aqueous solution (2) is 30 to 95%, and further 50 It is preferable to set it to -80%. The liquid passing speed and the liquid passing ratio when contacting the organic solvent solution (2) are the same as in the case of the organic solvent aqueous solution (1).

It is preferable to collect a fraction having a non-polymer catechin concentration of 0.1% by mass or more from the eluate contacted with the organic solvent aqueous solution.
Further, when non-polymer catechins are eluted from the synthetic adsorbent with an organic solvent aqueous solution, the concentration of non-polymer catechins in the leading portion of the elution collection fraction (the previous fraction with respect to the maximum value of the elution peak) is 0. It is preferable from the viewpoint of yield and removal of contaminants to collect a fraction of 1% by mass or more, further 0.2% by mass or more, particularly 0.4% by mass or more. The recovered fraction of the tailing portion of the eluted and recovered fraction (the fraction that is later with respect to the maximum value of the eluted peak) is the same as that of the leading portion.

  The synthetic adsorbent used in the present invention can be reused after the first adsorption-wash-elution operation. Specifically, as the regeneration treatment, an insoluble component such as caffeine adsorbed on the synthetic adsorbent is desorbed by passing an organic solvent aqueous solution such as ethanol. Alternatively, a method of passing and washing an alkaline aqueous solution such as sodium hydroxide and desorbing all the water-soluble components remaining on the synthetic adsorbent can be used. Further, cleaning with water vapor may be combined. The concentration of the organic solvent aqueous solution used in the regeneration treatment is preferably 50 to 95%, and the concentration of the alkaline aqueous solution is preferably 0.1 to 10% from the viewpoint of removing impurities remaining in the synthetic adsorbent.

  The purified green tea extract eluted with an organic solvent aqueous solution can be used as it is in the production of beverages, but it is preferable to contact with activated carbon in terms of color tone improvement and thickening improvement.

Examples of the raw material for the activated carbon include coconut shell, wood, and coal, with wood being preferred. Examples of the activated carbon activation method include a steam activation method, a gas activation method, and a chemical activation method, and the chemical activation method is preferable. For example, ZN-50, Y-10S, GS-1, GS-B (manufactured by Ajinomoto Fine Techno), Kuraray Coal GLC, Kuraray Coal PK-D, Kuraray Coal PW-D, Kuraray Coal GW, Kuraray Coal GA, Kuraray Coal GA-D, Kuraray Coal RP-15 (manufactured by Kuraray Chemical Co., Ltd.), white birch AW50, white birch A, white birch P, white birch KL, white birch M, white birch C, carborafine, WH2C (manufactured by Nippon Envirochemicals), GM130A, CW130A, CW130AR , CW350AR, GL130A, SG, SGA, SGP (manufactured by Phutamura Chemical), coconut, MAS mark, plum bee mark, plum bee F mark (manufactured by Taihei Chemical Sangyo), CPG, CAL, S80A (manufactured by Mitsubishi Chemical Calgon), etc. Product can be used.
From the viewpoint of improving the color tone of the product, reducing the amount of activated carbon used, and improving the recovery rate, the following are preferable as the activated carbon. The average pore diameter is preferably 0.5 to 10 nm (nanometer), more preferably 1.0 to 9.0 nm, and particularly preferably 2.0 to 8.0 nm. The pore volume is preferably 0.01 to 2.5 mL / g, more preferably 0.1 to 2.0 mL / g, and particularly preferably 0.5 to 1.7 mL / g. The specific surface area is 800~2000m ² / g, further 900~1600m ² / g, particularly preferably in the range of 1000~1500m ² / g. These physical property values are values based on the nitrogen adsorption method.

  The activated carbon is preferably added in an amount of 0.3 to 10 parts by weight, particularly 0.5 to 5 parts by weight, based on 100 parts by weight of the eluate based on the organic solvent aqueous solution. If the amount of activated carbon added is too small, the removal efficiency of caffeine and off-flavors will deteriorate, which is not preferable.

  The contact treatment between the eluate and activated carbon may be performed by any method such as a batch method or a continuous treatment using a column. In general, powdered activated carbon etc. is added and stirred, and after selectively adsorbing impurities such as caffeine, a method of obtaining a filtrate from which impurities have been removed by filtration operation, or a column packed with granular activated carbon etc. is used. For example, a method of selectively adsorbing impurities by continuous treatment is employed. It is good to carry out by a method such as continuous treatment using an activated carbon column.

  These adsorption-washing-elution operations are preferably performed at a temperature of 10 to 60 ° C, more preferably 20 to 40 ° C.

  The purified green tea extract with reduced nitrogen content, bitterness, sourness, thickness and hue obtained by the method of the present invention is particularly preferred when used for beverages such as sports drinks and isotonic beverages.

  The purified green tea extract obtained by the present invention contains 25 to 95% by mass of non-polymer catechins, further 40 to 95% by mass, more preferably 50 to 90% by mass, particularly 55 to 80% by mass in the solid content thereof. It is preferable to do this.

  Moreover, the ratio in the total non-polymer catechins of the gallate body which consists of catechin gallate, epicatechin gallate, gallocatechin gallate, and epigallocatechin gallate in the refined green tea extract obtained by this invention is 0-70 mass%. Further, it is preferably 0 to 50% by mass, and particularly preferably 0 to 40% by mass from the viewpoint of reducing the bitterness of non-polymer catechins.

  The concentration of caffeine in the purified green tea extract obtained in the present invention is such that caffeine / non-polymer catechins (mass ratio) = 0 to 0.15, more preferably 0 to 0.1 relative to non-polymer catechins. In particular, 0 to 0.05, particularly 0 to 0.035 is preferable in terms of improving taste.

  The concentration of gallic acid in the purified green tea extract obtained in the present invention is gallic acid / non-polymer catechins (mass ratio) = 0 to 0 with respect to non-polymer catechins from the viewpoint of reducing the sourness. 0.1, more preferably from 0 to 0.05, especially from 0 to 0.00028, and even more preferably from 0 to 0.00025.

  In addition, from the viewpoint of taste such as thickening of the purified green tea extract obtained in the present invention, nitrogen content / non-polymer catechins (mass%) = 0 to 0.26% by weight, and further 0.02 to 0.0. 25% by weight, particularly 0.03 to 0.24% by weight, is preferred.

  The purified green tea extract obtained by the present invention includes 25 to 95% by mass of non-polymer catechins in the solid content, 0 to 70% by mass of non-polymer catechin gallate body, gallic acid / non-polymer catechins ( (Mass ratio) 0 to 0.1, caffeine / non-polymer catechins (mass ratio) is 0 to 0.15, and nitrogen content / non-polymer catechins (mass%) is 0 to 0.00. What is 26 mass% is preferable at the point of taste improvement.

  The purified green tea extract obtained in the present invention can be used as it is. Further, the solvent may be removed by a method such as vacuum concentration or thin film concentration. When powder is desirable as the product form of the green tea extract, it can be pulverized by a method such as spray drying or freeze drying.

  The purified green tea extract obtained in the present invention can be blended into a container-packed beverage (green tea beverage, sports drink, isotonic beverage, etc.). Containers to be used are provided in ordinary forms such as molded containers (so-called PET bottles) mainly composed of polyethylene terephthalate, metal cans, paper containers combined with metal foil or plastic film, bottles, etc., as with general beverages. be able to. The term “packaged beverage” as used herein means a beverage that can be drunk without dilution.

  Moreover, said container-packed drink is manufactured on the sterilization conditions prescribed | regulated to the food hygiene law, for example, when it can heat-sterilize after filling a container like a metal can. For PET bottles and paper containers that cannot be sterilized by retort, sterilize under the same conditions as above, for example, after sterilizing at high temperature and short time with a plate heat exchanger, etc. The method is adopted. Moreover, you may mix | blend another component with the filled container under aseptic conditions.

(Measurement method of catechin, caffeine and gallic acid)
The sample solution was filtered with a filter (0.45 μm), and a high performance liquid chromatograph (model SCL-10AVP) manufactured by Shimadzu Corporation was used to pack an octadecyl group-introduced packed column L-column TM ODS (4.6 mmφ × 250 mm). : Chemical Substance Evaluation Research Organization) and a gradient method 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. .

(Measurement method of tannase activity)
Reagent A: pH 5.5 citrate buffer solution 50 mmol: 10.5 g of citric acid is dissolved in 800 mL of distilled water, adjusted to pH 5.5 with 1N NaOH solution, and diluted to 1000 mL.
Reagent B: 0.35 mass% substrate aqueous solution (tannic acid): 175 mg of tannic acid is dissolved in 50 mL of citrate buffer solution (reagent A).
Reagent C: 90 vol% ethanol solution.
Measuring method 1. Collect 1.0 mL of the substrate solution (reagent B) in a test tube and keep at 30 ° C. for 5 minutes.
2. Add 0.25 mL of sample solution and incubate at 30 ° C. for 15 minutes. In the blank solution, a citrate buffer solution (reagent A) is added instead of the sample solution.
3. Add 5.0 mL of ethanol solution (reagent C) to the sample solution and blank solution to stop the enzyme reaction.
4. Measure absorbance at 310 nm [sample: A _s , blank: A ₀ ].
The activity is calculated by the following formula.
Activity per volume (U / mL) = (A _s −A ₀ ) × 20.3 × 1.0 (mL) × 1.04 × df / (0.71 × 0.25 (mL) × 15 (min )) = ΔA × 7.93 × df
Activity per mass (U / g) = (U / mL) × 1 / C
20.3: μmol of tannic acid contained in 1.0 mL of the substrate solution (reagent B).
0.71: Change in absorbance after complete hydrolysis of 20.3 μmol of tannic acid under analytical conditions, 1.04: conversion factor, df: dilution factor, C: in sample (g / mL) Tannase concentration.

(Evaluation of color tone)
Using a spectrophotometer of HITACHI (model U-2001 type), measurement was performed by diluting with ion-exchanged water so that the concentration of non-polymer catechins in the sample was 0.180% by mass in a glass cell. The measurement wavelength of the spectrophotometer at the time of analysis was set to 400 nm.

(Nitrogen content)
Using a trace total nitrogen analyzer (decompression method) (model TN-100 Mitsubishi Chemical), dilute with ion-exchanged water so that the concentration of non-polymer catechins in the sample is 0.180% by mass in the vial. And measured.

(Evaluation of purified product)
The purified tea extract obtained in each example was diluted with deionized water so that the non-polymer catechins content was 0.175% [w / v], and 40 mL thereof was put into a 50 mL pressure-resistant glass container. I put it in. Thereto was added 0.1% by mass of Na ascorbate, the pH was adjusted to 6.4 with a 5% aqueous sodium bicarbonate solution, nitrogen substitution was performed, and the mixture was sterilized by heating in an autoclave at 121 ° C. for 10 minutes. Thereafter, the evaluation panel evaluated the bitterness, sourness, and thickness of the aftertaste by five evaluation panelists. The bitterness was evaluated by the quinine sulfate method.
As used herein, the term “boromi” refers to a flavor close to umami and an unpleasant complex sensation.

(Evaluation of bitterness by quinine sulfate method (equivalent concentration test method))
Quinine sulfate dihydrate was adjusted to a concentration corresponding to the bitterness intensity shown in Table 1. After tasting the evaluation sample, it was determined which sample of the standard bitterness solution had the same bitterness intensity. The bitterness intensity was confirmed by five evaluation panelists. (Reference: New edition Sensory Test Handbook, Nikkatsu Rensen Sensory Test Committee p448-449, Perception & Psychophysics, 5, 1696, 347-351)

Example 1
153 g of tannase-treated green tea extract is dissolved in 4591 g of ion-exchanged water to obtain “adsorption raw material liquid 1”. “Adsorption raw material liquid 1” contained 0.93 mass% of non-polymer catechins, and the gallate content of the non-polymer catechins composition was 29.9 mass%. Moreover, 0.17 mass% of caffeine and gallic acid / non-polymer catechins (mass ratio) were 0.122. The non-polymer catechins in the solid content of the tea extract was 30.4% by mass.
Next, 1186 mL of a synthetic adsorbent SP-70 (manufactured by Mitsubishi Chemical Corporation) was packed in a stainless steel column 1 (inner diameter 70 mm × height 480 mm, volume 1651 mL). In both columns, SV = 5 (h ⁻¹ ) and 95 (v / v) ethanol was passed through twice the volume (to the packing resin), and then water was passed through 20 times the volume (to the packing resin). Washed. Thereafter, 4744 g (4.0 times volume vs. synthetic adsorbent) of the obtained “adsorption raw material liquid 1” was passed through the column 1 at SV = 1 (h ⁻¹ ), the permeate was discarded, and “adsorption treatment resin 1 " Next, the synthetic adsorbent was washed with 1779 mL (1.5 times volume vs. synthetic adsorbent) of 0.01% by mass sodium hydroxide aqueous solution (pH 9.6) at SV = 1 (h ⁻¹ ). Discarded. Thereafter, 4151 mL (3.5 times volume vs. synthetic adsorbent) of 25% ethanol aqueous solution was flowed at SV = 1 (h ⁻¹ ) to elute non-polymer catechins, and among the leading fractions in the eluate The fraction having a non-polymer catechin concentration of 0.4% by weight or more was recovered to obtain 3865 g of “resin-treated product 1”. This extract contained 0.98% by mass of non-polymer catechins (turbidity 57 NTU), and the gallate content of the non-polymer catechins composition was 31.0% by mass. Further, caffeine was 0.16% by mass, gallic acid / non-polymer catechins (mass ratio) were 0.0003, and the color tone at 400 nm was 0.432. The non-polymer catechins in the solid content of the tea extract was 63.0% by mass.
Next, the solution eluted with 2372 mL (2 volumes vs. synthetic adsorbent) with 95% aqueous ethanol solution SV = 1 (h ⁻¹ ) on the synthetic adsorbent brought into contact with 25% ethanol aqueous solution was discarded.
Next, “resin-treated product 1” was subjected to concentration treatment under reduced pressure (2.6 kPa, 40 ° C.) with a non-polymer catechin concentration of 6.39 wt% (turbidity 187 NTU), and then 1.64 wt% catechin, A stainless steel column 2 (inner diameter: 22 mm × height: 190 mm, volume: 72.75 g) obtained by concentrating and diluting the solvent to 38% ethanol with 343.0 g of liquid and packed with 9.5 g (66.6 mL) of activated carbon (SGP manufactured by Phthamura Chemical). 2 mL), 72.2 mL (1.0 times volume to column volume) of ion-exchanged water, and ethanol was distilled off by concentration under reduced pressure (2.6 kPa, 40 ° C.). Green tea extract 1 "was obtained. The gallate content of the purified non-polymer catechin composition was 26.2% by mass. Further, 0% by mass of caffeine, gallic acid amount / non-polymer catechins (mass ratio) is 0.0002, color tone at 400 nm is 0.57, pH 4.7, nitrogen content / non-polymer catechins (mass%) ) Was 0.238%. In the evaluation of the purified product, there was little bitterness and sourness, and there was little little thickening.

Example 2
To the “adsorption treatment resin 1” obtained in Example 1, SV = 1 (h ⁻¹ ), 1779 mL (1.5 times volume vs. synthetic adsorbent) of 0.01 mass% aqueous sodium hydroxide solution (pH 9.6) ), The synthetic adsorbent was washed, and the liquid was discarded. Next, 4151 mL (3.5 times volume vs. synthetic adsorbent) of 25% ethanol aqueous solution at SV = 1 (h ⁻¹ ) was eluted to elute non-polymer catechins, and among the leading fractions in the eluate, Fractions having a polymer catechin concentration of 0.4% by weight or more were collected to obtain 3747 g of “resin-treated product 2”. This extract contained 1.00% by mass of non-polymer catechins (turbidity of 58 NTU), and the gallate content of the non-polymer catechins composition was 30.4% by mass. The caffeine content was 0.165% by mass, the gallic acid / non-polymer catechins (mass ratio) were 0.0003, and the color tone at 400 nm was 0.444. The non-polymer catechins in the solid content of the tea extract was 62.8% by mass.
Next, “resin-treated product 2” was subjected to concentration treatment under reduced pressure (2.6 kPa, 40 ° C.) with a non-polymer catechin concentration of 4.95 wt% (turbidity: 464 NTU), and then catechin 1.65 wt%, A stainless steel column 2 (inner diameter 22 mm × height 190 mm, volume 72.75 g) filled with 9.5 g (66.6 mL) of activated carbon (SGP manufactured by Futamura Chemical Co., Ltd.) was prepared by concentrating and diluting the solvent to 38% ethanol. 2 mL), 72.2 mL (1.0 times volume to column volume) of ion-exchanged water, and ethanol was distilled off by concentration under reduced pressure (2.6 kPa, 40 ° C.). Green tea extract 2 "was obtained. The gallate content of the purified non-polymer catechin composition was 26.6% by mass. Caffeine 0% by mass, gallic acid / non-polymer catechins (mass ratio) is 0.0002, color tone at 400 nm is 0.53, pH 4.6, nitrogen content / non-polymer catechins (mass%) Was 0.240%. In the evaluation of the purified product, there was little bitterness and sourness, and there was little little thickening.

Example 3
To the “adsorption treatment resin 1” obtained in Example 1, SV = 1 (h ⁻¹ ), 1779 mL (1.5 times volume vs. synthetic adsorbent) of 0.01 mass% aqueous sodium hydroxide solution (pH 9.6) ), The synthetic adsorbent was washed, and the liquid was discarded. Next, 4151 mL (3.5 times volume vs. synthetic adsorbent) of 25% ethanol aqueous solution at SV = 1 (h ⁻¹ ) was eluted to elute non-polymer catechins, and among the leading fractions in the eluate, The fraction having a polymer catechin concentration of 0.4% by weight or more was recovered to obtain 3622 g of “resin-treated product 3”. This extract contained 1.02% by mass of non-polymer catechins (turbidity 60 NTU), and the gallate content of the non-polymer catechins composition was 29.8% by mass. Further, 0.170% by mass of caffeine, gallic acid / non-polymer catechins (mass ratio) were 0.0003, and the color tone at 400 nm was 0.457. The non-polymer catechins in the solid content of the tea extract was 62.7% by mass.
Next, “resin-treated product 3” was subjected to concentration treatment under reduced pressure (2.6 kPa, 40 ° C.) with a non-polymer catechin concentration of 4.11% by weight (turbidity 473 NTU), and then 1.68% by weight of catechin, Stainless column 2 (inner diameter 22 mm × height 190 mm, volume 72.2 mL) filled with 9.4 g (65.6 mL) of activated carbon (SGP manufactured by Futamura Chemical Co., Ltd.) was obtained by concentrating and diluting the solvent to 38% ethanol. Then, 72.2 mL (1.0 times volume to column volume) of ion-exchanged water was passed through, and ethanol was distilled off by concentration under reduced pressure (2.6 kPa, 40 ° C.). Item 3 "was obtained. The gallate content of the purified non-polymer catechin composition was 27.3% by mass. Caffeine 0% by mass, gallic acid / non-polymer catechins (mass ratio) is 0.0002, color tone at 400 nm is 0.51, pH 4.6, nitrogen content / non-polymer catechins (mass%) Was 0.213%. In the evaluation of the purified product, there was little bitterness and sourness, and there was little thickness.

Comparative Example 1
The synthetic adsorbent was added to the “adsorption treatment resin 1” obtained in Example 1 with 1779 mL (1.5 times volume vs. synthetic adsorbent) ion-exchanged water (pH 6.0) at SV = 1 (h ⁻¹ ). Washed and discarded the solution. Next, 4151 mL (3.5 times volume vs. synthetic adsorbent) of 25% ethanol aqueous solution at SV = 1 (h ⁻¹ ) was eluted to elute non-polymer catechins to obtain 3987 g of “resin-treated product 4”. . This extract contained 0.96% by mass of non-polymer catechins, and the gallate content of the non-polymer catechins composition was 31.8% by mass. In addition, caffeine was 0.167% by mass, gallic acid / non-polymer catechins (mass ratio) were 0.0003, and the color tone at 400 nm was 0.653. The non-polymer catechins in the solid content of the tea extract was 62.9% by mass.
Next, “resin-treated product 4” was subjected to concentration treatment under reduced pressure (2.6 kPa, 40 ° C.), and then 611 g of liquid obtained by adjusting the concentration and dilution of catechin to 1.63% by weight and ethanol to 38%, The solution was passed through a stainless steel column 3 (inner diameter 22 mm × height 300 mm, volume 114 mL) packed with 15.5 g (108.2 mL) of activated carbon (SGP manufactured by Phthamura Chemical Co.), and an additional 114 mL (1.0 times volume vs. column volume). After passing ion-exchanged water, ethanol was distilled off by concentration under reduced pressure (2.6 kPa, 40 ° C.) to obtain “Purified Green Tea Extract 4”. The gallate content of the purified non-polymer catechin composition was 26.1% by mass. Moreover, 0 mass% of caffeine, gallic acid / non-polymer catechin (mass ratio) is 0.0004, color tone at 400 nm is 0.06, nitrogen content / non-polymer catechin (mass%) is 0.260. %Met. In the evaluation of the purified product, there was little bitterness and sourness, but there was a lot of thickening.

Comparative Example 2
The synthetic adsorbent was added to the “adsorption treatment resin 1” obtained in Example 1 with 1779 mL (1.5 times volume vs. synthetic adsorbent) ion-exchanged water (pH 6.0) at SV = 1 (h ⁻¹ ). Washed and discarded the solution. Next, 4151 mL (3.5 times volume vs. synthetic adsorbent) of 25% ethanol aqueous solution at SV = 1 (h ⁻¹ ) was eluted to elute non-polymer catechins, and among the leading fractions in the eluate, Fractions having a polymer catechin concentration of 0.3% by weight or more were collected to obtain 3850 g of “Resin-treated product 5”. This extract contained 1.00% by mass of non-polymer catechins, and the gallate content of the non-polymer catechins composition was 29.9% by mass. Further, 0.160% by mass of caffeine, gallic acid / non-polymer catechins (mass ratio) were 0.0004, and the color tone at 400 nm was 0.306. The non-polymer catechins in the solid content of the tea extract was 64.3% by mass.
Next, “resin-treated product 5” was concentrated under reduced pressure (2.6 kPa, 40 ° C.), non-polymer catechins were concentrated at 4.11 wt% (turbidity 473 NTU), and then catechin 1 A stainless steel column 4 (inner diameter 22 mm × height 210 mm, packed with 9.7 g (67.8 mL) of activated carbon (SGP manufactured by Futamura Chemical Co., Ltd.), a liquid of 347 g, which was concentrated and diluted to 38% ethanol and the solvent was adjusted to 38% ethanol. After passing 79.8 mL (1.0 times volume to column volume) of ion-exchanged water, ethanol was distilled off by concentration under reduced pressure (2.6 kPa, 40 ° C.). , “Purified green tea extract 5” was obtained. The gallate content of the purified non-polymer catechin composition was 25.1% by mass. Caffeine 0 mass%, gallic acid / non-polymer catechins (mass ratio) is 0.0004, pH 4.6, color tone at 400 nm is 0.04, nitrogen content / non-polymer catechins (mass%) Was 0.245%. In the evaluation of the purified product, there was little bitterness and sourness, but there was a little thickness.

Comparative Example 3
1779 mL (1.5 times volume vs. synthetic adsorbent) in which 3.6 g of citric acid was dissolved in ion exchange water at SV = 1 (h ⁻¹ ) in “adsorption treatment resin 1” obtained in Example 1 The synthetic adsorbent was washed with an acidic aqueous solution, and the liquid was discarded. Next, 4151 mL (3.5 times volume vs. synthetic adsorbent) of a 25% aqueous ethanol solution at SV = 1 (h ⁻¹ ) was eluted to elute non-polymer catechins, and the eluate eluted the concentration of non-polymer catechins. Among the leading fractions in the liquid, a fraction having a non-polymer catechin concentration of 0.5% by weight or more was recovered to obtain 3635 g of “resin-treated product 6”. This extract contained 0.98% by mass of non-polymer catechins, and the gallate content of the non-polymer catechins composition was 32.5% by mass. The caffeine content was 0.198% by mass, the gallic acid / non-polymer catechins (mass ratio) were 0.0006, and the color tone at 400 nm was 0.382. The non-polymer catechins in the solid content of the tea extract was 58.9% by mass.
Next, “resin-treated product 6” was subjected to concentration treatment under reduced pressure (2.6 kPa, 40 ° C.), and then 307 g of liquid obtained by concentrating and diluting the catechin to 1.71 wt% and the solvent to 38% ethanol, The solution was passed through a stainless steel column 4 (inner diameter 22 mm × height 210 mm, volume 79.8 mL) packed with 8.6 g (59.8 mL) of activated carbon (SGP manufactured by Futamura Chemical Co., Ltd.), and an additional 79.8 mL (1.0 times volume pair) Column volume) ion-exchanged water was passed through, and then ethanol was distilled off by concentration under reduced pressure (2.6 kPa, 40 ° C.) to obtain “purified green tea extract 6”. The gallate content of the purified non-polymer catechin composition was 28.9% by mass. Moreover, caffeine 0 mass%, gallic acid / non-polymer catechins (mass ratio) is 0.0003, pH 2.7, color tone at 400 nm is 0.02, nitrogen content / non-polymer catechins (mass%) Was 0.263%. In the evaluation of the purified product, there was little bitterness, but there was much acidity and thickness.

Comparative Example 4
4.7 g of green tea extract is dissolved in 140.9 g of ion exchange water to obtain “adsorption raw material liquid 2”. “Adsorption raw material liquid 2” contained 0.98% by mass of non-polymer catechins, and the gallate content of the non-polymer catechins composition was 51.2% by mass. In addition, 0.18% by mass of caffeine and gallic acid / non-polymer catechins (mass ratio) were 0.007. The amount of non-polymer catechins in the solid content of the tea extract was 31.8% by mass.
Subsequently, 36 mL of synthetic adsorbent SP-70 (manufactured by Mitsubishi Chemical Corporation) was packed in a stainless steel column 5 (inner diameter 22 mm × height 96 mm, volume 36.5 mL). Both columns were previously washed with SV = 5 (h ^-1 ) and 95 (v / v) ethanol through 2 volumes (to the packing resin) and then water with 20 volumes (to the packing resin). did. Thereafter, 145.6 g (4.0 times volume to synthetic adsorbent) of the obtained “adsorption raw material liquid 2” was passed through the stainless steel column 5 at SV = 1 (h ⁻¹ ), and the permeate was discarded. Treatment resin 2 "was obtained. Subsequently, the synthetic adsorbent was washed with 36.5 mL (1.0 volume vs. stainless steel column) of ion exchange water (pH 6.0) at SV = 2 (h ⁻¹ ), and the liquid was discarded. Thereafter, 489.5 mL (15 times volume vs. synthetic adsorbent) of 0.1 wt% NaOH aqueous solution at SV = 5 (h ⁻¹ ) was eluted to elute non-polymer catechins, and 483 g of “resin-treated product 7” Got. This extract contained 0.16% by mass of non-polymer catechins, and the gallate content of the non-polymer catechins composition was 54.5% by mass. The content of caffeine was 0% by mass, but the pH was 9.6, the color tone was poor (black), and catechin was decomposed.

  The green tea extracts described in Examples 1 to 3 and Comparative Examples 1 to 4 were sterilized based on the Food Sanitation Law and evaluated for bitterness and thickness. The results are also shown in Table 2.

  In Examples 1 to 3, a green tea extract having a reduced caffeine concentration, a reduced hue and nitrogen content, and reduced bitterness, sourness, and thickness is obtained. In Comparative Examples 1 and 2, the resin washing liquid is ion-exchanged. It is water, has a high nitrogen content, and is thick. In Comparative Example 3, the resin cleaning solution is an acidic aqueous solution, so even if the elution and recovery solution is subjected to fraction removal, the nitrogen content is high and thick. In Comparative Example 4, the color tone was bad and the decomposition of non-polymer catechins was observed in Comparative Example 4 because a large amount of high-concentration alkaline solution was passed.

Claims (7)

  A green tea extract is adsorbed on a synthetic adsorbent, a basic aqueous solution is brought into contact with the synthetic adsorbent, the synthetic adsorbent is washed, and then an organic solvent aqueous solution is brought into contact with the synthetic resin adsorbent. Of purified green tea extract that elutes fruits The process according to claim 1 Symbol placement pH of the basic aqueous solution is 7 to 11 for cleaning the synthetic adsorbent. The method according to claim 1 or 2 , wherein the concentration of the organic solvent aqueous solution for eluting non-polymer catechins is 5 to 50%. The method according to any one of claims 1 to 3 , wherein the organic solvent aqueous solution is an ethanol aqueous solution. The manufacturing method of any one of Claims 1-4 which collect | recovers fractions with a non-polymer catechin density | concentration of 0.1 mass% or more among the eluates which contacted the organic-solvent aqueous solution. The production method according to any one of claims 1 to 5 , wherein the eluate of the organic solvent aqueous solution is further brought into contact with activated carbon. The method according to any one of claims 1 to 6 , wherein the green tea extract is obtained by hydrolysis treatment.