JP4244230B2 - Method for producing purified tea extract - Google Patents
Method for producing purified tea extract Download PDFInfo
- Publication number
- JP4244230B2 JP4244230B2 JP2006056350A JP2006056350A JP4244230B2 JP 4244230 B2 JP4244230 B2 JP 4244230B2 JP 2006056350 A JP2006056350 A JP 2006056350A JP 2006056350 A JP2006056350 A JP 2006056350A JP 4244230 B2 JP4244230 B2 JP 4244230B2
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- JP
- Japan
- Prior art keywords
- mass
- tea extract
- polymer
- polymer catechins
- synthetic adsorbent
- Prior art date
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- 238000004519 manufacturing process Methods 0.000 title claims description 8
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- 150000001765 catechin Chemical class 0.000 claims description 69
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 claims description 60
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- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 description 1
- 235000010378 sodium ascorbate Nutrition 0.000 description 1
- 229960005055 sodium ascorbate Drugs 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 description 1
- 235000019614 sour taste Nutrition 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000003440 styrenes Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229920001864 tannin Polymers 0.000 description 1
- 235000018553 tannin Nutrition 0.000 description 1
- 239000001648 tannin Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
Description
本発明は、非重合体カテキンガレート体率を低減し、カフェイン量を低減し、没食子酸を低減することにより呈味が改善された精製茶抽出物の製造方法に関する。 The present invention relates to a method for producing a purified tea extract having improved taste by reducing the percentage of non-polymer catechin gallate, reducing the amount of caffeine, and reducing gallic acid.
カテキンの生理効果としてはαアミラーゼ活性阻害作用などが報告されている（例えば、特許文献１参照）。このような生理効果を発現させるためには、成人一日あたりお茶を４〜５杯飲むことが必要であることから、より簡便に大量のカテキンを摂取するため、飲料にカテキンを高濃度配合する方法が望まれていた。 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, it is necessary to drink 4 to 5 cups of tea per day for an adult. Therefore, in order to ingest a large amount of catechin more easily, a high concentration of catechin is added to the beverage. A method was desired.
この方法の一つとして、緑茶抽出物の濃縮物（特許文献２）などの茶抽出物を利用して、カテキンを飲料に溶解状態で添加する方法が用いられている。しかしながら、この方法によりカテキンを高濃度に配合する対象となる飲料の種によっては、例えば紅茶抽出液や炭酸飲料にカテキンを添加する場合など、カフェイン及び緑茶由来の苦渋みの残存が飲料の商品価値を大きく損ねることがわかっている。 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 to be 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.
紅茶等の発酵茶抽出液に対してタンナーゼ処理を行い、低温冷却時の懸濁、即ちテイークリーム形成を抑制できることは古くから知られていた。又、特許文献３に見られる、ガレート体カテキンにタンナーゼ処理を行い、一部又は全部を没食子酸とすることにより、カテキン類と没食子酸との混合物を得る方法によれば、苦味の原因となるガレート体カテキン類を低減することができる。また、茶抽出物から、カフェイン等の夾雑物を取り除く方法としては、吸着法（特許文献４〜６）、抽出法（特許文献７）等が知られている。
しかしながら、タンナーゼ処理により得られるカテキン類と没食子酸の混合物は、酸味・エグ味が発生するという問題があった。 However, the mixture of catechins and gallic acid obtained by tannase treatment has a problem that sourness / egg taste occurs.
従って、本発明の課題は、非重合体カテキンガレート体率を低減し、かつ呈味の改善された精製茶抽出物の製造法を提供することにある。 Therefore, the subject of this invention is providing the manufacturing method of the refined tea extract which reduced the non-polymer catechin gallate body rate and improved the taste.
本発明者らは、茶抽出物中の非重合体カテキン類精製処理を検討した結果、加水分解処理して非重合体カテキンガレート体を遊離型非重合体カテキン類と没食子酸に分解し、非重合体カテキンガレート体率を低減させる第一の工程、更に、加水分解処理を行った後の茶抽出物を合成吸着剤に通液し、一旦、合成吸着剤に吸着させ、次いで、吸着した非重合体カテキン類を塩基性水溶液により溶出させる第二の工程を行うことにより、非重合体カテキンガレート体率が低減し、カフェイン含量及び、副生する没食子酸を低減させた呈味の改善された茶抽出物が得られることを見出した。 As a result of examining the purification treatment of non-polymer catechins in the tea extract, the present inventors hydrolyzed and decomposed the non-polymer catechin gallate into free non-polymer catechins and gallic acid. The first step of reducing the polymer catechin gallate content rate, and further, the tea extract after the hydrolysis treatment is passed through the synthetic adsorbent, once adsorbed to the synthetic adsorbent, and then adsorbed By performing the second step of eluting the polymer catechins with a basic aqueous solution, the non-polymer catechin gallate body ratio is reduced, and the taste is improved by reducing the caffeine content and by-product gallic acid. It was found that a tea extract was obtained.
本発明は、茶抽出物を加水分解後、合成吸着剤に吸着させ、合成吸着剤を有機溶媒濃度０〜２０質量％の有機溶媒水溶液で洗浄し、次いで合成吸着剤に塩基性水溶液を接触させて非重合体カテキン類を溶出させる、下記（ａ）〜（ｄ）を有する精製茶抽出物の製造方法を提供するものである。
（ａ）固形分中の非重合体カテキン類２５〜９５質量％
（ｂ）非重合体カテキンガレート体０〜７０質量％
（ｃ）没食子酸と非重合体カテキン類との比率が０〜０．１
（ｄ）カフェインと非重合体カテキン類の比率が０〜０．１５
In the present invention, after the tea extract is hydrolyzed, it is adsorbed on a synthetic adsorbent, the synthetic adsorbent is washed with an organic solvent aqueous solution having an organic solvent concentration of 0 to 20% by mass, and then the basic adsorbent is brought into contact with the synthetic adsorbent. The present invention provides a method for producing a purified tea extract having the following (a) to (d) for eluting non-polymer catechins.
(A ) Non-polymer catechins in solid content of 25 to 95% by mass
( B) Non-polymer catechin gallate body 0-70 mass%
( C) The ratio of gallic acid to non-polymer catechins is 0 to 0.1
( D) The ratio of caffeine to non-polymer catechins is 0 to 0.15
本発明により、処理前と比較して、非重合体カテキンガレート体率が低減され、カフェイン含量が低く、更に加水分解処理の際に副生する没食子酸が大幅に低減され、呈味の改善された精製茶抽出物を得ることができる。 According to the present invention, the ratio of non-polymer catechin gallate is reduced, the caffeine content is low, and gallic acid by-produced during the hydrolysis treatment is greatly reduced compared to before treatment, thereby improving taste. Purified tea extract can be obtained.
本発明で非重合体カテキン類とは、カテキン、ガロカテキン、カテキンガレート、ガロカテキンガレートなどの非エピ体カテキン及びエピカテキン、エピガロカテキン、エピカテキンガレート、エピガロカテキンガレートなどのエピ体カテキンをあわせての総称である。 In the present invention, the non-polymer catechins include non-epi 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.
本発明で非重合体カテキンガレート体とは、カテキンガレート、ガロカテキンガレート、エピカテキンガレート、エピガロカテキンガレートなどをあわせての総称である。 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.
本発明で用いる茶抽出物としては、緑茶、紅茶、烏龍茶等の茶葉から得られた抽出液が挙げられる。その他のカフェイン含有植物由来、例えばコーヒー等のカフェイン含有抽出物と茶抽出液の混合物等も用いることができる。使用する茶葉としては、より具体的には、Camellia属、例えばC.sinensis、C.assamica及びやぶきた種又はそれらの雑種等から得られる茶葉から製茶された茶葉が挙げられる。製茶された茶葉には、煎茶、番茶、玉露、てん茶、釜炒り茶等の緑茶類がある。また、超臨界状態の二酸化炭素接触処理を施した茶葉を用いてもよい。 Examples of the tea extract used in the present invention include extracts obtained from tea leaves such as green tea, black tea and oolong tea. A mixture of a caffeine-containing extract such as coffee and a tea extract derived from other caffeine-containing plants can also be used. 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.
茶葉からの抽出は、抽出溶媒として水又は水溶性有機溶媒又はそれらの混合物を使用し、攪拌抽出等により行われる。抽出の際、水又は水溶性有機溶媒又はそれらの混合物にあらかじめアスコルビン酸ナトリウム等の有機酸塩類又は有機酸を添加してもよい。また、煮沸脱気や窒素ガス等の不活性ガスを通気して溶存酸素を除去しつつ、いわゆる非酸化的雰囲気下で抽出する方法を併用してもよい。このようにして得られた抽出液は、そのままでも、乾燥、濃縮しても本発明に使用できる。茶抽出物の形態としては、液体、スラリー、半固体、固体の状態が挙げられる。 Extraction from 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 tea extract include liquid, slurry, semi-solid, and solid state.
本発明に使用する茶抽出物には、茶葉から抽出した抽出液を使用する代わりに、茶抽出物の濃縮物を水又は有機溶媒に溶解又は水又は有機溶媒に希釈して用いても、茶葉からの抽出液と茶抽出物の濃縮物とを併用してもよい。
ここで、茶抽出物の濃縮物とは、茶葉から熱水又は水溶性有機溶媒により抽出された抽出物を濃縮したものであり、例えば、特開昭５９−２１９３８４号公報、特開平４−２０５８９号公報、特開平５−２６０９０７号公報、特開平５−３０６２７９号公報等に記載されている方法により調製したものをいう。具体的には、緑茶抽出物として、市販の東京フードテクノ社製「ポリフェノン」、伊藤園社製「テアフラン」、太陽化学社製「サンフェノン」等の粗カテキン製剤を固体の緑茶抽出物として用いることもできる。
For the tea extract used in the present invention, instead of using an extract extracted from tea leaves, a concentrate of 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 tea, and the concentrate of a tea extract.
Here, the concentrate of tea extract is obtained by concentrating an extract extracted from tea leaves with hot water or a water-soluble organic solvent. For example, JP-A-59-219384 and JP-A-4-20589. No. 5, 260-907, JP-A-5-306279, and the like. Specifically, as a green tea extract, a commercially available crude catechin preparation such as “Polyphenone” manufactured by Tokyo Food Techno Co., “Theafuran” manufactured by ITO EN Co., Ltd., “Sunphenon” manufactured by Taiyo Kagaku Co., Ltd. may be used as a solid green tea extract. it can.
本発明においては、まず茶抽出物を加水分解処理する。当該加水分解処理により、非重合体カテキン類中のガレート体率が低下する。加水分解による非重合体カテキン類中の非重合体カテキンガレート体の濃度減少は、呈味改善の点から５質量％以上、更に７質量％以上、特に１０質量％以上が好ましい。加水分解の方法は、酵素類による処理、酸処理、アルカリ処理等により行なわれる。酵素類としては、タンナーゼ活性を有する酵素、菌体又は培養液、酸としては、塩酸、硫酸、リン酸、アルカリとしては苛性ソーダなどが好ましい。その中でも反応制御の点から酵素類での加水分解が好ましい。ここでタンナーゼ活性を有するとは、タンニンを分解する活性を有することを意味し、本活性を有すれば任意の酵素、菌体、培養液が使用できる。 In the present invention, the tea extract is first hydrolyzed. By the hydrolysis treatment, the gallate content in the non-polymer catechins decreases. 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.
具体的には、タンナーゼ活性を有する酵素として市販品では、ペクチナーゼＰＬアマノ（天野エンザイム社製）、ヘミセルラーゼアマノ９０（天野エンザイム社製）、タンナーゼＫＴＦＨ（キッコーマン社製）等が利用できる。その中でもタンナーゼが好ましい。例えば、アスペルギルス属、ペニシリウム属、リゾプス属のタンナーゼ生産菌を培養して得られるタンナーゼが挙げられる。このうちアスペルギルス オリーゼ由来のものが好ましい。
タンナーゼ活性を有する菌体とは、タンナーゼ活性を有する酵素を産生することができる菌体であり、麹菌等があげられる。例えば、アスペルギルス属、ペニシリウム属等が挙げられ、このうちアスペルギルス オリーゼが好ましい。
タンナーゼ活性を有する培養液とは、アスペルギルス属、ペニシリウム属、リゾプス属のタンナーゼ生産菌を培養して得られる培養液である。好ましくは、タンニン酸を唯一の炭素源として培養して得られる培養液があげることができ、精製品であっても未精製なものであっても用いることができる。
風味劣化の抑制及び生産性の点から加水分解を極力短時間で終了する際は、酵素又は培養液を利用することが好ましい。
本発明で使用するタンナーゼ活性を有する酵素又は培養液は、５００〜１００，０００Ｕ／ｇの酵素活性を有することが好ましく、５００Ｕ／ｇ以下であると工業的に限られた時間内で処理するためには多量の酵素が必要となり、１００，０００Ｕ／ｇ以上であると酵素反応速度が速すぎる為、反応系を制御することが困難となる。ここで1Ｕｎｉｔは３０℃の水中においてタンニン酸に含まれるエステル結合を１マイクロモル加水分解する酵素量を示す。
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. For example, Aspergillus genus, Penicillium genus and the like can be mentioned, among which Aspergillus oryzae is preferable.
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 used, and it can be used as a purified product or an unpurified product.
When hydrolysis is completed in a short time as much as possible from the viewpoint of suppression of flavor deterioration and productivity, 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 when it is 500 U / g or less. Requires a large amount of enzyme, and if it is 100,000 U / g or more, the enzyme reaction rate is too high, and it becomes difficult to control the reaction system. 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.
タンナーゼ活性を有する酵素及び培養液による処理を行うときの非重合体カテキン濃度は、好ましくは０．１〜２２質量％、更に好ましくは０．１〜１５質量％、特に好ましくは０．５〜１０質量％、殊更好ましくは０．５〜３質量％である。０．１質量％未満ではこの後の合成吸着剤への吸着時に吸着量が低下し、２２質量％を超えると、加水分解処理に長時間を要し、生産性及び茶抽出物の味の点から好ましくない。
呈味を改善した非重合体カテキンガレート体率を得るため、茶抽出物中の非重合体カテキン類に対して酵素又は培養液を０．０１〜１０質量％の範囲になるように添加することが好ましい。酵素失活の工程を含め、上記加水分解処理を工業的に最適な酵素反応時間である２時間以内で終了させるためには、酵素又は培養液濃度が０．０１〜７質量％、更に０．０３〜５質量％であることが好ましい。
緑茶抽出物中の非重合体カテキン類に対してタンナーゼ活性を有する酵素又は培養液を、好ましくは１〜３００Ｕｎｉｔ／ｇ−非重合体カテキン、更に好ましくは３〜２００Ｕｎｉｔ／ｇ−非重合体カテキン、特に好ましくは５〜１５０Ｕｎｉｔ／ｇ−非重合体カテキンになるように添加する。
酵素又は培養液による処理の温度は、最適な酵素活性が得られる０〜７０℃が好ましく、更に好ましくは０〜６０℃、特に好ましくは５〜５０℃である。
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 the amount is less than 0.1% by mass, the amount of adsorption decreases during the subsequent adsorption to the synthetic adsorbent. If the amount exceeds 22% by mass, the hydrolysis process takes a long time, and productivity and the taste of the tea extract are required. Is not preferable.
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 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 having a tannase activity with respect to non-polymer catechins in a green tea extract or a culture solution, preferably 1 to 300 Unit / g-non-polymer catechin, more preferably 3 to 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.
酵素又は培養液での加水分解反応を終了させるには、酵素活性を失活させる必要がある。酵素失活は、加熱ことにより達成される。酵素失活温度は、７０〜１００℃が好ましく、７０℃未満では酵素を短時間で充分に失活することが困難であるため加水分解反応が進行し、非重合体カテキンガレート体率の範囲内で酵素反応を停止することができない。又、失活温度に到達してからの保持時間が１０秒程度以下であると酵素活性を充分に失活することが困難であるため、酵素反応が進行し、また、２０分以上の保持時間では、非重合体カテキン類の非エピメリ化が起こる場合があり好ましくない。
酵素反応の失活方法は、バッチ式もしくはプレート型熱交換機のような連続式で加熱を行うことで停止することができる。又、タンナーゼ処理の失活終了後、遠心分離などの操作により茶抽出物を清澄化することができる。
In order to terminate the hydrolysis reaction with the enzyme or the culture solution, it is necessary to deactivate the enzyme activity. Enzyme inactivation is achieved by heating. The enzyme deactivation temperature is preferably 70 to 100 ° C. If the enzyme deactivation temperature is less than 70 ° C., it is difficult to deactivate the enzyme sufficiently in a short time, so that the hydrolysis reaction proceeds and the non-polymer catechin gallate ratio is within the range. The enzyme reaction cannot be stopped. Further, if the retention time after reaching the deactivation temperature is about 10 seconds or less, it is difficult to sufficiently deactivate the enzyme activity, so that the enzyme reaction proceeds, and the retention time of 20 minutes or more Then, non-epimerization of non-polymer catechins may occur, which is not preferable.
The deactivation method of the enzyme reaction can be stopped by heating in a batch system or a continuous system such as a plate heat exchanger. In addition, after the inactivation of the tannase treatment, the tea extract can be clarified by an operation such as centrifugation.
菌体として例えば麹菌を利用する場合は、非重合体カテキンの濃度が、好ましくは０．１〜２２質量％、更に好ましくは０．１〜１５質量％、特に好ましくは０．５〜１５質量％である茶抽出物に麹菌を入れ加水分解処理を行なう。麹菌は、その種類等により大幅に異なるが、茶抽出物中の非重合体カテキン類に対して通常０．５質量％〜１０質量％の範囲内、特に1．０質量％〜５質量％の範囲内で添加される。温度条件としては、４５℃〜７０℃、更に５０〜６０℃が好ましい。醗酵時間は通常１２時間〜２０日間、更に１日〜１０日間で行われることが好ましい。麹菌の酵素活性の失活は、酵素又は培養液での加水分解反応を終了させる時と同様である。 When, for example, gonococcus is used as the microbial cell, the concentration of non-polymer catechin 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. A koji mold is added to the tea extract, which is then hydrolyzed. Neisseria gonorrhoeae varies greatly depending on its type and the like, but is usually in the range of 0.5 mass% to 10 mass%, particularly 1.0 mass% to 5 mass% with respect to the non-polymer catechins in the tea extract It is added within the range. As temperature conditions, 45 to 70 degreeC, Furthermore, 50 to 60 degreeC is preferable. The fermentation time is usually 12 hours to 20 days, and 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.
加水分解後の茶抽出物を合成吸着剤に吸着させ、次いで塩基性水溶液を接触させて非重合体カテキン類を溶出させる。当該合成吸着剤処理により、カフェイン及び没食子酸が低減できる。
吸着後、塩基性水溶液を接触させる前に、合成吸着剤を洗浄し、合成吸着剤中の没食子酸や不純物を除去するのが好ましい。
合成吸着剤は、一般に不溶性の三次元架橋構造ポリマーでイオン交換基のような官能基を実質的に持たないものである。好ましくは、イオン交換基が１meq／ｇ未満のものを用いることができる。本発明に用いる合成吸着剤としては、その母体がスチレン系、例えばアンバーライトＸＡＤ４、ＸＡＤ１６ＨＰ、ＸＡＤ１１８０、ＸＡＤ２０００、（供給元：米国ローム＆ハース社）；ダイヤイオンＨＰ２０、ＨＰ２１（三菱化学社製）；セパビーズＳＰ８５０、ＳＰ８２５、ＳＰ７００、ＳＰ７０（三菱化学社製）；ＶＰＯＣ１０６２（Ｂａｙｅｒ社製）、臭素原子を核置換して吸着力を強めた修飾スチレン系、例えばセパビーズＳＰ２０５、ＳＰ２０６、ＳＰ２０７（三菱化学社製）、メタクリル系、例えばダイヤイオンＨＰ１ＭＧ、ＨＰ２ＭＧ（三菱化学社製）、フェノール系、例えばアンバーライトＸＡＤ７６１（ロームアンドハース社製）、アクリル系、例えばアンバーライトＸＡＤ７ＨＰ（ロームアンドハース社製）、ポリビニル系、例えばＴＯＹＯＰＥＡＲＬ、ＨＷ-４０Ｃ（東ソー社製）、デキストラン系、例えばＳＥＰＨＡＤＥＸ、ＬＨ-20（ファルマシア社製）等が使用できる。
合成吸着剤としては、その母体がスチレン系、メタクリル系、アクリル系、ポリビニル系が好ましく、特にスチレン系がカテキンとカフェインとの分離性の点から好ましい。
The tea extract after hydrolysis is adsorbed on a synthetic adsorbent, and then contacted with a basic aqueous solution to elute non-polymer catechins. 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 (supplier: Rohm & Haas, USA); Diaion HP20, HP21 (manufactured by Mitsubishi Chemical); Sepabeads SP850, SP825, SP700, SP70 (manufactured by Mitsubishi Chemical); VPOC1062 (manufactured by Bayer), modified styrene based on nuclear substitution of bromine atoms to enhance the adsorptive power, such as sepabeads SP205, SP206, SP207 (manufactured by Mitsubishi Chemical) ), Methacrylic such as Diaion HP1MG, HP2MG (Mitsubishi Chemical), phenolic such as Amberlite XAD761 (Rohm and Haas), acrylic such as Amberlite XAD7HP (Rohm and Haas), Polybi Le system, e.g. TOYOPEARL, HW-40C (manufactured by Tosoh Corp.), dextran based, for example SEPHADEX, LH-20 (Pharmacia) or 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.
加水分解処理後の茶抽出物を合成吸着剤に吸着させる手段としては、加水分解処理後の茶抽出物に合成吸着剤を添加、撹拌し吸着後、ろ過操作により合成吸着剤を回収するバッチ方法又は合成吸着剤を充填したカラムを用いて連続処理により吸着処理を行なうカラム方法が採用されるが、生産性の点からカラムによる連続処理方法が好ましい。
合成吸着剤が充填されたカラムは、予めＳＶ（空間速度）＝０．５〜１０［ｈ-1］、合成吸着剤に対する通液倍数として２〜１０［ｖ／ｖ］の通液条件で９５ｖｏｌ％エタノール水溶液による洗浄を行い、合成吸着剤の原料モノマーや原料モノマー中の不純物等を除去するのが好ましい。そして、その後ＳＶ＝０．５〜１０［ｈ-1］、合成吸着剤に対する通液倍数として１〜６０［ｖ／ｖ］の通液条件により水洗を行い、エタノールを除去して合成吸着剤の含液を水系に置換する方法により非重合体カテキン類の吸着能が向上する。
As a means of adsorbing the hydrolyzed tea extract to the synthetic adsorbent, the synthetic adsorbent is added to the hydrolyzed tea extract, stirred and adsorbed, and then the synthetic adsorbent is recovered by filtration. Alternatively, a column method in which adsorption treatment is performed by continuous treatment using a column filled with a synthetic adsorbent is adopted, but a continuous treatment method using a column is preferable from the viewpoint of productivity.
The column packed with the synthetic adsorbent is preliminarily SV (space velocity) = 0.5 to 10 [h −1 ], and 95 vol under a liquid passing condition of 2 to 10 [v / v] as the liquid passing ratio with respect to the synthetic adsorbent. It is preferable to remove the raw material monomer of the synthetic adsorbent, impurities in the raw material monomer, etc. by washing with a% ethanol aqueous solution. 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.
加水分解処理後の茶抽出物を合成吸着剤に吸着させる手段としては、合成吸着剤が充填されたカラムに当該茶抽出物を通液するのが好ましい。茶抽出物を合成吸着剤の充填したカラムに通液する条件としては、ＳＶ（空間速度）＝０．５〜１０［ｈ-1］の通液速度で、合成吸着剤に対する通液倍数として０．５〜２０［ｖ／ｖ］で通液するのが好ましい。１０［ｈ-1］以上の通液速度や２０［ｖ／ｖ］以上の通液量であると非重合体カテキン類の吸着が不充分又は不安定となる場合がある。
更に、加水分解処理後の茶抽出物を合成吸着剤に吸着させた後に洗浄する場合は、洗浄に使用する水溶液としては、カテキンの回収率の点からｐH７以下が好ましく、水溶性有機溶媒との混合系においても使用することができる。水溶性有機溶媒としては、アセトン、メタノール、エタノールなどが挙げられ、食品への使用の観点から、エタノールが好ましい。含有する有機溶媒の濃度は、０〜２０質量％、好ましくは０〜１０質量％、より好ましくは０〜５質量％がカテキンの回収率の点から好ましい。
ＳＶ（空間速度）＝０．５〜１０[ｈ-1]の通液速度で、合成吸着剤に対する通液倍数として１〜１０[ｖ／ｖ]で、合成吸着剤に付着した没食子酸や不純物を除去するのが好ましい。更にＳＶ＝０．５〜５[ｈ-1] の通液速度で、通液倍数として１〜５[ｖ／ｖ] で洗浄することが没食子酸や不純物の除去及び非重合体カテキン類の回収率の点から好ましい。
As a means for adsorbing the tea extract after the hydrolysis treatment to the synthetic adsorbent, it is preferable to pass the tea extract through a column filled with the synthetic adsorbent. The conditions for passing the 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.
Further, when the tea extract after hydrolysis treatment is adsorbed on a synthetic adsorbent and then washed, the aqueous solution used for washing is preferably pH 7 or less from the viewpoint of the recovery rate of catechin, It can also be used in mixed systems. 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.
SV (space velocity) = 0.5 to 10 [h-1], and gallic acid and impurities adhering to the synthetic adsorbent at a flow rate of 1 to 10 [v / v] with respect to the synthetic adsorbent. Is preferably removed. Furthermore, it is possible to remove gallic acid and impurities and to recover non-polymer catechins by washing at a flow rate of SV = 0.5 to 5 [h-1] and a flow rate of 1 to 5 [v / v]. It is preferable in terms of rate.
非重合体カテキン類の溶出に用いる塩基性水溶液としては、アルカリ金属塩、アルカリ土類のアルカリ水溶液、好ましくは、ナトリウム系のアルカリ性水溶液、例えば水酸化ナトリウム水溶液、炭酸ナトリウム水溶液等を好適に用いることができる。また、アルカリ性水溶液のｐＨは７〜１４の範囲が好ましい。非重合体カテキン類回収率の点から９〜１３．８、特に１０〜１３．５が好ましい。ｐＨ７〜１４のナトリウム系水溶液としては、４％以下の水酸化ナトリウム水溶液、１Ｎ−炭酸ナトリウム水溶液等が挙げられる。塩基性水溶液に、水溶性有機溶媒が含まれていてもよい。有機溶媒の濃度としては、カフェインとカテキンの分離性の点から０〜９０質量％の範囲が好ましく、０〜５０質量％がより好ましく、０〜２０質量％が更に好ましい。 As the basic aqueous solution used for elution of non-polymer catechins, 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 are preferably used. Can do. The pH of the alkaline aqueous solution is preferably in the range of 7-14. From the point of non-polymer catechin recovery, 9 to 13.8, particularly 10 to 13.5 is preferable. Examples of the sodium-based aqueous solution having a pH of 7 to 14 include a 4% or less sodium hydroxide aqueous solution and a 1N-sodium carbonate aqueous solution. The basic aqueous solution may contain a water-soluble organic solvent. As a density | concentration of an organic solvent, the range of 0-90 mass% is preferable from the point of the separability of caffeine and catechin, 0-50 mass% is more preferable, 0-20 mass% is still more preferable.
溶出工程においては、溶出に用いる塩基性水溶液として互いにｐＨが異なる２種以上の塩基性水溶液を用い、これら塩基性水溶液をｐＨが低い順に合成吸着剤に接触させることができる。それぞれのｐＨ区分で異なる非重合体カテキン類や他の成分を脱着することができる。 In the elution step, two or more basic aqueous solutions having different pH values are used as the basic aqueous solution used for elution, and these basic aqueous solutions can be brought into contact with the synthetic adsorbent in order of decreasing pH. Different non-polymer catechins and other components can be desorbed in each pH category.
本発明で使用される合成吸着剤は本発明実施後に再使用できる。再生処理としては、具体的には、エタノールのような有機溶媒を通液し合成吸着剤上に吸着したカフェイン等の不溶分を脱着させる。又は水酸化ナトリウムのようなアルカリ水溶液を通液・洗浄し、合成吸着剤上に残存する水溶性成分をすべて脱着させるなどの方法が挙げられる。更に水蒸気による洗浄を組み合わせても良い。 The synthetic adsorbent used in the present invention can be reused after the practice of the present invention. Specifically, as the regeneration treatment, an insoluble component such as caffeine adsorbed on the synthetic adsorbent is passed through an organic solvent 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.
非重合体カテキン類の溶出液は、塩基性水溶液で溶出したため塩基性であり、非重合カテキン類の安定性の観点から、溶出液のｐＨを７以下に調整することが好ましい。具体的には酸による中和、電気透析によるアルカリ金属イオンの除去、又はカチオン交換樹脂によるアルカリ金属イオンの除去が利用できる。イオン交換樹脂としては特にＨ型のカチオン交換樹脂を用いるのが好ましい。プロセスの簡便性からイオン交換樹脂によるｐＨ調整が好ましい。カチオン交換樹脂としては、具体的には、アンバーライト２００ＣＴ、ＩＲ１２０Ｂ、ＩＲ１２４、ＩＲ１１８、ダイヤイオンＳＫ１Ｂ、ＳＫ１ＢＨ、ＳＫ１０２、ＰＫ２０８、ＰＫ２１２等を用いることができる。 The eluate of non-polymer catechins is basic because it is eluted with a basic aqueous solution, and the pH of the eluate is preferably adjusted to 7 or less from the viewpoint of the stability of the non-polymer catechins. Specifically, neutralization with an acid, removal of alkali metal ions by electrodialysis, or removal of alkali metal ions by a cation exchange resin can be used. As the ion exchange resin, it is particularly preferable to use an H-type cation exchange resin. From the simplicity of the process, pH adjustment with an ion exchange resin is preferred. Specifically, Amberlite 200CT, IR120B, IR124, IR118, Diaion SK1B, SK1BH, SK102, PK208, PK212, etc. can be used as the cation exchange resin.
更に非重合体カテキン類の溶出液は、必要に応じて濃縮して後の工程に付すことができる。 Further, the eluate of non-polymer catechins can be concentrated as necessary and subjected to a subsequent step.
得られた非重合体カテキン類の溶出液が懸濁する場合は、除濁することが好ましい。除濁の具体的な操作としては、ろ過及び／又は遠心分離処理により固形分と水溶性部分とを固液分離することが挙げられる。 When the eluate of the obtained non-polymer catechins is suspended, it is preferably turbid. Specific operation of turbidity includes solid-liquid separation of solid content and water-soluble part by filtration and / or centrifugation.
本発明によって得られる精製茶抽出物は、その固形分中に、非重合体カテキン類を２５〜９５質量％含有するが、４０〜９５質量％、更に５０〜９０質量％、特に５５〜８０質量％含有するのが好ましい。 The purified tea extract obtained by the present invention contains 25 to 95% by mass of non-polymer catechins in its solid content, but it is 40 to 95% by mass, more preferably 50 to 90% by mass, especially 55 to 80% by mass. % Content is preferable.
また、本発明により得られる精製茶抽出物中のカテキンガレート、エピカテキンガレート、ガロカテキンガレート及びエピガロカテキンガレートからなるガレート体の全非重合体カテキン類中での割合は、０〜７０質量％、更に０〜５０質量％であるのが好ましく、特に０〜４０質量％であるのが、非重合体カテキン類の苦味低減の点で好ましい。 In the purified tea extract obtained by the present invention, the proportion of gallate bodies composed of catechin gallate, epicatechin gallate, gallocatechin gallate and epigallocatechin gallate in all non-polymer catechins is 0 to 70% by 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.
本発明で得られる精製茶抽出物中のカフェイン濃度は、非重合体カテキン類に対して、カフェイン／非重合体カテキン類（質量比）＝０〜０．１５、更に０〜０．１、特に０〜０．０５、殊更に０〜０．０３５であるのが呈味改善の点で好ましい。 The caffeine concentration in the purified 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 with respect to non-polymer catechins. In particular, 0 to 0.05, particularly 0 to 0.035 is preferable in terms of improving taste.
また、本発明で得られる精製茶抽出物中の没食子酸濃度は、苦味、酸味等の呈味の点から、非重合体カテキン類に対して、没食子酸／非重合体カテキン類（質量比）＝０〜０．１、更に０〜０．０７、特に０〜０．０５が好ましい。 The concentration of gallic acid in the purified tea extract obtained in the present invention is gallic acid / non-polymer catechins (mass ratio) with respect to non-polymer catechins from the viewpoint of taste such as bitterness and sourness. = 0 to 0.1, more preferably 0 to 0.07, and particularly preferably 0 to 0.05.
本発明で得られる精製茶抽出物としては、固形分中に非重合体カテキン類を２５〜９５質量％、非重合体カテキンガレート体率０〜７０質量％、没食子酸／非重合体カテキン類（質量比）０〜０．１であり、カフェイン／非重合体カテキン類（質量比）が０〜０．１５であるものが、呈味改善の点で好ましい。 The purified tea extract obtained in the present invention contains 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 ( A mass ratio of 0 to 0.1 and a caffeine / non-polymer catechin (mass ratio) of 0 to 0.15 are preferred in terms of taste improvement.
飲料の色調の安定性向上のため、非重合体カテキン類の溶出液を脱色することが好ましい。具体的な脱色操作としては、茶抽出物を水又は有機溶媒との混合液に分散また溶解し、活性炭又は／及び、活性白土又は／及び酸性白土に接触させることにより、脱色することができる。 In order to improve the stability of the color tone of the beverage, it is preferable to decolorize the eluate of non-polymer catechins. As a specific decolorization operation, the tea extract is dispersed or dissolved in a mixed solution with water or an organic solvent, and decolorized by contacting with activated carbon or / and activated clay or / and acidic clay.
本発明で得られた精製茶抽出物はそのままで使用できる。また、減圧濃縮、薄膜濃縮などの方法により溶媒を除去してもよい。また茶抽出物の製品形態として粉体が望ましい場合は、噴霧乾燥や凍結乾燥等の方法により粉体化できる。 The purified 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 desired as the product form of the tea extract, it can be pulverized by a method such as spray drying or freeze drying.
本発明で得られた精製茶抽出物は容器詰飲料に配合できる。使用される容器は一般の飲料と同様にポリエチレンテレフタレートを主成分とする成形容器（いわゆるＰＥＴボトル）、金属缶、金属箔やプラスチックフィルムと複合された紙容器、瓶などの通常の形態で提供することができる。ここでいう容器詰飲料とは希釈せずに飲用できるものをいう。 The purified tea extract obtained in the present invention can be blended in a container-packed beverage. 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.
また上記の容器詰飲料は、例えば、金属缶のように容器に充填後、加熱殺菌できる場合にあっては食品衛生法に定められた殺菌条件で製造される。ＰＥＴボトル、紙容器のようにレトルト殺菌できないものについては、あらかじめ上記と同等の殺菌条件、例えばプレート式熱交換器などで高温短時間殺菌後、一定の温度迄冷却して容器に充填する等の方法が採用される。また無菌下で、充填された容器に別の成分を配合して充填してもよい。 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.
（カテキン、カフェイン及び没食子酸の測定法）
試料溶液をフィルター（０．４５μｍ）で濾過し、島津製作所製、高速液体クロマトグラフ（型式ＳＣＬ−１０ＡＶＰ）を用い、オクタデシル基導入液体クロマトグラフ用パックドカラムＬ−カラムＴＭ ＯＤＳ（４．６ｍｍφ×２５０ｍｍ：財団法人 化学物質評価研究機構製）を装着し、カラム温度３５℃でグラジエント法で行った。カテキン類の標準品としては、三井農林製のものを使用し、検量線法で定量した。移動相Ａ液は酢酸を０．１ｍｏｌ／Ｌ含有の蒸留水溶液、Ｂ液は酢酸を０．１ｍｏｌ／Ｌ含有のアセトニトリル溶液とし、試料注入量は２０μＬ、ＵＶ検出器波長は２８０ｎｍの条件で行った。
(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. .
（タンナーゼ活性の測定法）
試薬Ａ：ｐＨ５．５クエン酸緩衝溶液５０ｍｍｏｌ：蒸留水８００ｍＬにクエン酸１０．５ｇを溶解し、１ＮのＮａＯＨ溶液でｐＨ５．５に調整し、１０００ｍＬに希釈する。
試薬Ｂ：０．３５質量％基質水溶液（タンニン酸）：５０ｍＬクエン酸緩衝溶液（試薬Ａ）にタンニン酸１７５ｍｇを溶解する。
試薬Ｃ：９０ｖｏｌ％エタノール溶液。
測定方法
１．試験管に基質溶液（試薬Ｂ）を１．０ｍＬ採取し、３０℃で５分間保つ。
２．試料溶液０．２５ｍＬ添加し、１５分間３０℃で培養する。ブランク溶液は、試料溶液の代わりにクエン酸緩衝溶液（試薬Ａ）を加える。
３．酵素反応を停止するため試料溶液とブランク溶液に５．０ｍＬのエタノール溶液（試薬Ｃ）を加える。
４．３１０ｎｍの吸光度を測定する［試料：Ａs、ブランク：Ａ0］。
次の計算式により活性を計算する。
体積当たりの活性（Ｕ／ｍＬ）＝（Ａs−Ａ0）×２０．３×１．０（ｍＬ）×１．０４×ｄｆ／（０．７１×０．２５（ｍＬ）×１５（ｍｉｎ））＝ΔＡ×７．９３×ｄｆ
質量当たりの活性（Ｕ／ｇ）＝（Ｕ／ｍＬ）×１／Ｃ
２０．３：基質溶液（試薬Ｂ）の１．０ｍＬ中に含まれるタンニン酸のμｍｏｌ。
０．７１：分析条件下での２０．３μｍｏｌのタンニン酸が完全に加水分解した後の吸光度の変化量、１．０４：換算係数、ｄｆ：希釈係数、Ｃ：サンプル（ｇ／ｍＬ）中のタンナーゼ濃度。
(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.
殺菌後の風味評価
各実施例で得られた茶抽出物をカテキン含有率が０．１７５％［ｗ／ｖ］となるように脱イオン水で希釈し、その４０ｍＬを５０ｍＬの耐圧製ガラス容器に入れた。そこにアスコルビン酸Ｎａを０．１質量％添加し、５％重炭酸Ｎａ水溶液でｐＨを６．４に調整し、窒素置換を行い、オートクレーブで１２１℃、１０分間加熱滅菌した。その後、評価パネラー５名によって緑茶由来の異味・異臭が感じられないか確認を行った。
Flavor evaluation after sterilization The tea extract obtained in each example was diluted with deionized water so that the catechin 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. Then, it was confirmed by the evaluation panelists whether or not the taste and odor derived from green tea were felt.
硫酸キニーネ法（等価濃度試験法）による苦味評価
硫酸キニーネ２水和物を表１に記載の苦味強度に対応した濃度に調整する。評価サンプルを試飲した後、標準苦味溶液のどのサンプルと苦味の強さが等しいか判断する。評価パネラー５名によって苦味強度の確認を行った（参考文献：新版官能検査ハンドブック 日科技連官能検査委員会p448-449、Perception & Psychophysics,5,1696,347-351）。
Evaluation of bitterness by quinine sulfate method (equivalent concentration test method) Quinine sulfate dihydrate is adjusted to a concentration corresponding to the bitterness intensity shown in Table 1. After tasting the evaluation sample, it is determined which sample of the standard bitterness solution has the same bitterness intensity. The evaluation panelists confirmed the bitterness intensity (reference: new edition sensory test handbook, Nikkagi Technical Sensory Test Committee p448-449, Perception & Psychophysics, 5,1696,347-351).
実施例１
緑茶葉（中国雲南省、大葉種）１．８ｋｇに９０℃の熱水２７ｋｇを添加し、３０分間攪拌バッチ抽出したのち、１００メッシュ金網で粗濾過、遠心分離操作後、２号濾紙による濾過を行い、「緑茶抽出液」２０．４ｋｇ（ｐＨ５．３）を得た。（緑茶抽出液の非重合体カテキン類濃度＝０．９６質量％、緑茶抽出液のガレート体率＝６９．５質量％、カフェイン＝０．２４質量％、没食子酸＝０．０１質量％）
この緑茶抽出液を温度２５℃に設定し、タンナーゼ（キッコーマン社製タンナーゼＫＴＦＨ、５００Ｕ／ｇ）を緑茶抽出液に対して３００ｐｐｍとなる濃度で添加し、８５分間保持し、ガレート体率５２．４質量％になったところで、９０℃に溶液を加熱して、２分間保持し酵素を失活させ、反応を止めた（ｐＨ４．８、タンナーゼ処理液（１））。
次いで、ステンレスカラム１（内径１１０ｍｍ×高さ２３０ｍｍ、容積２１８５ｍＬ）に充填した合成吸着剤ＳＰ−７０（三菱化学（株）製）２０４８ｍＬを、予めＳＶ＝５（ｈ-1）で９５（ｖ／ｖ）エタノール８１９２ｍＬによる洗浄を行い、次いでＳＶ＝５（ｈ-1）で２０４８０ｍＬの水で洗浄した。ステンレスカラム２（内径３８ｍｍ×高さ７７０ｍｍ、容積８７３ｍＬ）に充填したイオン交換樹脂ＳＫ１ＢＨ（三菱化学（株）製）８５２ｍＬを、予めＳＶ＝５（ｈ-1）で９５（ｖ／ｖ）エタノール３４０８ｍＬによる洗浄を行い、次いでＳＶ＝５（ｈ-1）で８５２０ｍＬの水で洗浄した。その後、タンナーゼ処理液（１）８１９２ｇ（４倍容積対合成吸着剤）をＳＶ＝１（ｈ-1）でカラム１に通液し透過液は廃棄した。次いでＳＶ＝２（ｈ-1）で２０４８ｍＬ（１倍容積対合成吸着剤）の水で洗浄した。水洗後、０．１質量％水酸化ナトリウム水溶液（ｐＨ１２．５）をＳＶ＝５（ｈ-1）で３０７２０ｍＬ通液し（１５倍容積対合成吸着剤）カテキン溶出液を得た。溶出液は連続でステンレスカラム２に通液し、脱イオンを行い、非重合体カテキン類組成物２８２２２ｇ（ｐＨ３．０）を得た。この組成物中には非重合体カテキン類０．２４質量％が含まれており、タンナーゼ処理液（１）からの非重合体カテキン類の回収率は９２．９％、非重合体カテキン類組成物のガレート体率は５５．３質量％であった。又、カフェイン０質量％、没食子酸量０．００１質量％であった。茶抽出物の固形分中の非重合体カテキン類６２．４質量％であった。
Example 1
After adding 27 kg of 90 ° C hot water to 1.8 kg of green tea leaves (Yunnan Province, China) and extracting with 30-minute stirring batch, coarse filtration with a 100-mesh wire mesh, centrifugal separation, and filtration with No. 2 filter paper And 20.4 kg of “green tea extract” (pH 5.3) was obtained. (Non-polymer catechin concentration of green tea extract = 0.96% by mass, gallate body ratio of green tea extract = 69.5% by mass, caffeine = 0.24% by mass, gallic acid = 0.01% by mass)
This green tea extract was set at a temperature of 25 ° C., tannase (Tannase KTFH, 500 U / g manufactured by Kikkoman) was added at a concentration of 300 ppm with respect to the green tea extract, held for 85 minutes, and a gallate body ratio of 52.4. When the mass was reached, the solution was heated to 90 ° C. and held for 2 minutes to inactivate the enzyme to stop the reaction (pH 4.8, tannase treatment solution (1)).
Next, 2048 mL of a synthetic adsorbent SP-70 (manufactured by Mitsubishi Chemical Corporation) packed in a stainless steel column 1 (inner diameter 110 mm × height 230 mm, volume 2185 mL) was preliminarily set to 95 (v / v) at SV = 5 (h −1 ). v) Washing with 8192 mL of ethanol followed by 20480 mL of water with SV = 5 (h −1 ). Ion exchange resin SK1BH (manufactured by Mitsubishi Chemical Corporation) 852 mL packed in stainless steel column 2 (inner diameter 38 mm × height 770 mm, volume 873 mL) was preliminarily set to SV = 5 (h −1 ) and 95 (v / v) ethanol 3408 mL. And then with 8520 mL water at SV = 5 (h −1 ). Thereafter, 8192 g of tannase treatment solution (1) (4 volumes vs. synthetic adsorbent) was passed through column 1 at SV = 1 (h −1 ), and the permeate was discarded. It was then washed with 2048 mL (1 volume vs. synthetic adsorbent) of water at SV = 2 (h −1 ). After washing with water, 30720 mL of 0.1 mass% aqueous sodium hydroxide solution (pH 12.5) was passed through with SV = 5 (h −1 ) (15 times volume vs. synthetic adsorbent) to obtain a catechin eluate. The eluate was continuously passed through the stainless steel column 2 and deionized to obtain 28222 g (pH 3.0) of a non-polymer catechin composition. This composition contains 0.24% by mass of non-polymer catechins, the recovery rate of non-polymer catechins from the tannase treatment liquid (1) is 92.9%, and the composition of non-polymer catechins The gallate content of the product was 55.3% by mass. The amount of caffeine was 0% by mass and the amount of gallic acid was 0.001% by mass. The non-polymer catechins in the solid content of the tea extract was 62.4% by mass.
比較例１
実施例１と同様にタンナーゼ処理を行ない、合成吸着剤への通液及び溶出は行わなかった。
Comparative Example 1
The tannase treatment was performed in the same manner as in Example 1, and the liquid was not passed through and eluted from the synthetic adsorbent.
比較例２
タンナーゼ処理を行わなかった以外は、実施例１と全く同様に行った。
Comparative Example 2
The procedure was the same as in Example 1 except that tannase treatment was not performed.
実施例２
（１）緑茶葉（ケニア産、大葉種）３ｋｇに８８℃の熱水４５ｋｇを添加し、６０分間攪拌バッチ抽出したのち、１００メッシュ金網で粗ろ過後、抽出液中の微粉を除去する為に遠心分離操作を行い、「緑茶抽出液」３７．２ｋｇ（ｐＨ５．４）を得た。（緑茶抽出液中の非重合体カテキン類濃度＝０．８９質量％、緑茶抽出液のガレート体率＝５２．３質量％、カフェイン０．１７質量％）
この緑茶抽出液を温度１５℃に保持し、タンナーゼ（キッコーマン社製タンナーゼＫＴＦＨ、５００Ｕ／ｇ）を緑茶抽出液に対して４３０ｐｐｍとなる濃度で添加し、５５分間保持し、ガレート体率３０．５質量％になったところで、９０℃に溶液を加熱して、２分間保持し酵素を失活させ、反応を止めた（ｐＨ５．１）。次いで減圧濃縮にて７０℃、６．７ｋｐａでＢｒｉｘ濃度２０％まで濃縮処理を行い、更に噴霧乾燥して粉末状の「タンナーゼ処理した緑茶抽出物」０．９ｋｇを得た。得られた緑茶抽出物は非重合体カテキン類含有量２７．８質量％、非重合体カテキンガレート体率３０．３質量％、カフェイン含有量６．７４質量％、没食子酸３．５８質量％であった。「タンナーゼ処理した緑茶抽出物」１０ｇを、脱イオン水３００ｇに２５℃で３０分間攪拌溶解した（タンナーゼ処理液（２）。
次いで、ステンレスカラム３（内径２２ｍｍ×高さ９６ｍｍ、容積３６．５ｍＬ）に充填した合成吸着剤ＳＰ−７０（三菱化学（株）製）３６．１ｍＬを、予め実施例1と同様の方法で洗浄し、ガラスカラム（内径１６ｍｍ×高さ８０ｍｍ、容積１６．１ｍＬ）に充填したイオン交換樹脂ＳＫ１ＢＨ（三菱化学（株）製）１４．７ｍＬを、予め実施例1と同様の方法で洗浄した。その後、得られたタンナーゼ処理液（２）１４４．４ｇ（４倍容積対合成吸着剤）をＳＶ＝１（ｈ-1）でカラム１に通液し透過液は廃棄した。次いでＳＶ＝２（ｈ-1）で３６．１ｍＬ（１倍容積対合成吸着剤）の水で洗浄した。水洗後、０．１質量％水酸化ナトリウム水溶液（ｐＨ１２．４）をＳＶ＝５（ｈ-1）で５６１．８ｍＬ通液した（１５倍容積対合成吸着剤）。溶出液は連続でガラスカラムに通液して、脱イオンを行い、非重合体カテキン類組成物５５２ｇ（ｐＨ２．７）を得た。この抽出物中には非重合体カテキン類０．２１質量％が含まれており、タンナーゼ処理液（２）からの非重合体カテキン類の回収率は９０．３％、非重合体カテキン類組成物のガレート体率は３２．５質量％、であった。又、カフェイン０質量％、没食子酸量０．００２質量％であった。茶抽出物の固形分中の非重合体カテキン類６４．７質量％であった。
Example 2
(1) Add 45kg of hot water at 88 ° C to 3kg of green tea leaves (Kenya, large leaf seeds), extract with stirring batch for 60 minutes, and after coarse filtration with 100 mesh wire netting, to remove fine powder in the extract Centrifugation was performed to obtain 37.2 kg (pH 5.4) of “green tea extract”. (Non-polymer catechin concentration in green tea extract = 0.89 mass%, gallate body ratio of green tea extract = 52.3 mass%, caffeine 0.17 mass%)
This green tea extract was kept at a temperature of 15 ° C., tannase (Tannase KTFH, manufactured by Kikkoman Corp., 500 U / g) was added at a concentration of 430 ppm with respect to the green tea extract, kept for 55 minutes, and a gallate body ratio of 30.5 When the mass was reached, the solution was heated to 90 ° C. and held for 2 minutes to deactivate the enzyme, and the reaction was stopped (pH 5.1). Next, concentration was performed under reduced pressure at 70 ° C. and 6.7 kpa to a Brix concentration of 20%, followed by spray drying to obtain 0.9 kg of a powdered “tannase-treated green tea extract”. The resulting green tea extract had a non-polymer catechin content of 27.8% by mass, a non-polymer catechin gallate content of 30.3% by mass, a caffeine content of 6.74% by mass, and a gallic acid of 3.58% by mass. Met. 10 g of “tannase-treated green tea extract” was dissolved in 300 g of deionized water with stirring at 25 ° C. for 30 minutes (tannase treatment solution (2).
Next, 36.1 mL of the synthetic adsorbent SP-70 (Mitsubishi Chemical Corporation) packed in the stainless steel column 3 (inner diameter 22 mm × height 96 mm, volume 36.5 mL) was previously washed in the same manner as in Example 1. Then, 14.7 mL of ion exchange resin SK1BH (manufactured by Mitsubishi Chemical Corporation) packed in a glass column (inner diameter 16 mm × height 80 mm, volume 16.1 mL) was previously washed in the same manner as in Example 1. Thereafter, 144.4 g of the tannase treatment liquid (2) obtained (4 times volume to the synthetic adsorbent) was passed through the column 1 at SV = 1 (h −1 ), and the permeate was discarded. It was then washed with 36.1 mL (1 volume vs. synthetic adsorbent) of water at SV = 2 (h −1 ). After washing with water, 561.8 mL of 0.1 mass% sodium hydroxide aqueous solution (pH 12.4) was passed at SV = 5 (h −1 ) (15 times volume vs. synthetic adsorbent). The eluate was continuously passed through a glass column and deionized to obtain 552 g (pH 2.7) of a non-polymer catechin composition. This extract contains 0.21% by mass of non-polymer catechins, the recovery rate of non-polymer catechins from the tannase treatment liquid (2) is 90.3%, and the composition of non-polymer catechins The gallate content of the product was 32.5% by mass. The amount of caffeine was 0% by mass and the amount of gallic acid was 0.002% by mass. The non-polymer catechins in the solid content of the tea extract was 64.7% by mass.
実施例３
粗カテキン製剤（三井農林製 非重合体カテキン類濃度＝３２．０質量％、非重合体カテキンガレート体率＝５２．０質量％、カフェイン＝５．５１質量％、没食子酸＝０．１７質量％）１０ｇを、脱イオン水３００ｇに２５℃で３０分間攪拌溶解し茶抽出物溶解液（ｐＨ５．３）を得た。次いで、タンナーゼ（キッコーマン社製タンナーゼＫＴＦＨ、５００Ｕ／ｇ）を茶抽出物溶解液に対して５００ｐｐｍとなる濃度で添加し、１５℃、１２０分間保持し、ガレート体率４質量％になったところで、９０℃に溶液を加熱して、２分間保持し酵素を失活させ、反応を止めた（ｐＨ４．２、タンナーゼ処理液（３））。
原料が上記タンナーゼ処理液以外は、実施例２と同様のカラム及び操作条件にて精製を行い、非重合体カテキン類組成物５３４．２ｇ（ｐＨ３．７）を得た。この抽出物中には非重合体カテキン類０．１８質量％が含まれており、タンナーゼ処理液（３）からの非重合体カテキン類の回収率は９１．３％、非重合体カテキン類組成物のガレート体率は６．０質量％、であった。又、カフェイン０質量％、没食子酸量０．００４質量％であった。茶抽出物の固形分中の非重合体カテキン類５７．６質量％であった。
Example 3
Crude catechin preparation (Mitsui Norin non-polymer catechin concentration = 32.0 mass%, non-polymer catechin gallate body ratio = 52.0 mass%, caffeine = 5.51 mass%, gallic acid = 0.17 mass %) 10 g was stirred and dissolved in 300 g of deionized water at 25 ° C. for 30 minutes to obtain a tea extract solution (pH 5.3). Next, tannase (Kikkoman Tannase KTFH, 500 U / g) was added at a concentration of 500 ppm with respect to the tea extract solution, held at 15 ° C. for 120 minutes, and when the gallate content was 4% by mass, The solution was heated to 90 ° C. and held for 2 minutes to inactivate the enzyme to stop the reaction (pH 4.2, tannase treatment solution (3)).
The raw material was purified using the same column and operating conditions as in Example 2 except that the tannase treatment solution was used to obtain 534.2 g (pH 3.7) of a non-polymer catechin composition. This extract contains 0.18% by mass of non-polymer catechins, the recovery rate of non-polymer catechins from the tannase treatment liquid (3) is 91.3%, and the composition of non-polymer catechins The gallate content of the product was 6.0% by mass. The amount of caffeine was 0% by mass and the amount of gallic acid was 0.004% by mass. The non-polymer catechins in the solid content of the tea extract was 57.6% by mass.
実施例４
ステンレスカラム４（内径６０ｍｍ×高さ３６０ｍｍ、容積１０１７ｍＬ）に充填した合成吸着剤ＳＰ−２０７（三菱化学（株）製）１００４ｍＬを、予めＳＶ＝ ５（ｈ-1）で９５（ｖ／ｖ）エタノール４０１６ｍＬによる洗浄を行い、次いでＳＶ＝５（ｈ-1）で１００４０ｍＬの水で洗浄した。ステンレスカラム５（内径３８ｍｍ×高さ１２００ｍｍ、１３６０容積ｍＬ）に充填したイオン交換樹脂ＳＫ１ＢＨ（三菱化学（株）製）１３３８ｍＬを、予めＳＶ＝５（ｈ-1）で９５（ｖ／ｖ）エタノール５３５２ｍＬによる洗浄を行い、次いでＳＶ＝５（ｈ-1）で１３３８０ｍＬの水で洗浄した。実施例１のタンナーゼ処理液４０１６ｇ（４倍容積対合成吸着剤）をＳＶ＝１（ｈ-1）でステンレスカラム４に通液し透過液は廃棄した。次いでＳＶ＝２（ｈ-1）で１００４ｍ（１倍容積対合成吸着剤）の水で洗浄した。水洗後、１質量％水酸化ナトリウム水溶液（ｐＨ１４．０）をＳＶ＝５（ｈ-1）で５０２０ｍＬ通液した（５倍容積対合成吸着剤）。溶出液は連続でステンレスカラム５に通液して、脱イオンを行い、非重合体カテキン類組成物４９７５．５ｇ（ｐＨ４．１）を得た。この抽出物中には非重合体カテキン類０．６３質量％が含まれており、タンナーゼ処理液（１）からの非重合体カテキン類の回収率は８８．２％、非重合体カテキン類組成物のガレート体率は５８．３質量％、であった。又、カフェイン０質量％、没食子酸量０．００１質量％であった。茶抽出物の固形分中の非重合体カテキン類５９．３質量％であった。
Example 4
Synthetic adsorbent SP-207 (Mitsubishi Chemical Corporation) 1004 mL packed in a stainless steel column 4 (inner diameter 60 mm × height 360 mm, volume 1017 mL) was previously 95 (v / v) at SV = 5 (h −1 ). Washing was performed with 4016 mL of ethanol followed by 10040 mL of water with SV = 5 (h −1 ). Ion exchange resin SK1BH (manufactured by Mitsubishi Chemical Corporation) 1338 mL packed in a stainless steel column 5 (inner diameter 38 mm × height 1200 mm, 1360 volume mL) was previously added with 95 (v / v) ethanol at SV = 5 (h −1 ). Washing with 5352 mL was followed by washing with 13380 mL water at SV = 5 (h −1 ). 4016 g of tannase treatment solution of Example 1 (4 volumes vs. synthetic adsorbent) was passed through the stainless steel column 4 at SV = 1 (h −1 ), and the permeate was discarded. It was then washed with 1004 m (1 volume vs. synthetic adsorbent) of water at SV = 2 (h −1 ). After washing with water, 5020 mL of 1 mass% aqueous sodium hydroxide solution (pH 14.0) was passed at SV = 5 (h −1 ) (5 times volume vs. synthetic adsorbent). The eluate was continuously passed through the stainless steel column 5 and deionized to obtain 4975.5 g (pH 4.1) of a non-polymer catechin composition. This extract contains 0.63% by mass of non-polymer catechins, the recovery rate of non-polymer catechins from the tannase treatment liquid (1) is 88.2%, and the composition of non-polymer catechins The gallate content of the product was 58.3% by mass. The amount of caffeine was 0% by mass and the amount of gallic acid was 0.001% by mass. It was 59.3% by mass of non-polymer catechins in the solid content of the tea extract.
実施例５
実施例２のイオン交換樹脂ＳＫ１ＢＨの充填量を３．６ｍＬとした以外は実施例２と同様の操作を行なった。得られた抽出物は非重合体カテキン０．１７質量％が含まれており、タンナーゼ処理液（２）からの非重合体カテキン類の回収率は７８．２％、非重合体カテキン類組成物のガレート体率は３５．８質量％、であった。又、カフェイン０質量％、没食子酸量０．００１質量％であった。茶抽出物の固形分中の非重合体カテキン類４３．３質量％であった。
Example 5
The same operation as in Example 2 was performed except that the filling amount of the ion exchange resin SK1BH of Example 2 was 3.6 mL. The obtained extract contains 0.17% by mass of non-polymer catechin, the recovery rate of non-polymer catechins from the tannase treatment liquid (2) is 78.2%, and the composition of non-polymer catechins The gallate content was 35.8% by mass. The amount of caffeine was 0% by mass and the amount of gallic acid was 0.001% by mass. The amount of non-polymer catechins in the solid content of the tea extract was 43.3% by mass.
比較例３
ステンレスカラム４（内径６０ｍｍ×高さ３６０ｍｍ、容積１０１７ｍＬ）に充填した合成吸着剤ＳＰ−７０（三菱化学（株）製）８６０ｍＬを、予めＳＶ＝５（ｈ-1）で９５（ｖ／ｖ）エタノール３４４０ｍＬによる洗浄を行い、次いでＳＶ＝５（ｈ-1）で８６００ｍＬの水で洗浄した。実施例１のタンナーゼ処理液３４４０ｇ（４倍容積対合成吸着剤）をＳＶ＝１（ｈ-1）でステンレスカラム４に通液し透過液は廃棄した。
次いでＳＶ＝２（ｈ-1）で８６０ｍＬ（１倍容積対合成吸着剤）の水で洗浄した。水洗後、２０質量％エタノール水溶液をＳＶ＝２（ｈ-1）で５１６０ｍＬ通液した（６倍容積対合成吸着剤）。４０℃、２．７ｋＰａでエタノールを留去し、その後、水分量を調整して得られた抽出物は非重合体カテキン０．５０質量％が含まれており、タンナーゼ処理液（１）からの非重合体カテキン類の回収率は８４．４％、非重合体カテキン類組成物のガレート体率は４５．９質量％であった。又、カフェイン０．１質量％、没食子酸量０．００８質量％であった。茶抽出物の固形分中の非重合体カテキン類６０．８質量％であった。
Comparative Example 3
Synthetic adsorbent SP-70 (manufactured by Mitsubishi Chemical Co., Ltd.) 860 mL packed in a stainless steel column 4 (inner diameter 60 mm × height 360 mm, volume 1017 mL) was preliminarily set to 95 (v / v) at SV = 5 (h −1 ). Washing was performed with 3440 mL of ethanol, and then with 8600 mL of water at SV = 5 (h −1 ). 3440 g of the tannase treatment solution of Example 1 (4 volumes vs. synthetic adsorbent) was passed through the stainless steel column 4 at SV = 1 (h −1 ), and the permeate was discarded.
It was then washed with 860 mL (1 volume vs. synthetic adsorbent) of water at SV = 2 (h −1 ). After washing with water, 5160 mL of a 20% by mass aqueous ethanol solution was passed at SV = 2 (h −1 ) (6 volumes vs. synthetic adsorbent). The extract obtained by distilling ethanol off at 40 ° C. and 2.7 kPa and then adjusting the water content contained 0.50% by mass of non-polymer catechin, and was extracted from the tannase treatment solution (1). The recovery rate of non-polymer catechins was 84.4%, and the gallate content of the non-polymer catechins composition was 45.9% by mass. The amount of caffeine was 0.1% by mass and the amount of gallic acid was 0.008% by mass. The non-polymer catechins in the solid content of the tea extract was 60.8% by mass.
比較例４
酸性白土（ミズカエース＃６００、水澤化学社製）１００ｇを常温、３５０ｒ／ｍｉｎ攪拌条件下の９２．４質量％エタノール水溶液８００ｇ中に分散させ、約１０分間攪拌を行った後、（実施例２）で得られたタンナーゼ処理した緑茶抽出物２００ｇを投入し、室温のまま約３時間の攪拌を続けた（ｐＨ４．０）。その後、生成している沈殿及び酸性白土を２号ろ紙でろ過した。得られたろ液を活性炭（クラレコールＧＬＣ、クラレケミカル社製）３０ｇと接触させ、続けて０．２μｍメンブランフィルターによってろ過を行った。最後にイオン交換水２００ｇを添加して、４０℃、２．７ｋＰａでエタノールを留去し、その後、水分量を調整して「精製緑茶抽出物」を得た。得られた抽出物は非重合体カテキン２０．２質量％が含まれており、実施例２のタンナーゼ処理した緑茶抽出物からの非重合体カテキン類の回収率は６０．５％、非重合体カテキン類組成物のガレート体率は２９．３質量％、であった。又、カフェイン０．７３質量％、没食子酸量２．５６質量％であった。茶抽出物の固形分中の非重合体カテキン類５６．６質量％であった。
Comparative Example 4
100 g of acid clay (Mizuka Ace # 600, manufactured by Mizusawa Chemical Co., Ltd.) was dispersed in 800 g of a 92.4 mass% ethanol aqueous solution under stirring conditions at room temperature and 350 r / min. After stirring for about 10 minutes, (Example 2) 200 g of the tannase-treated green tea extract obtained in the above was added, and stirring was continued for about 3 hours at room temperature (pH 4.0). Thereafter, the generated precipitate and acid clay were filtered with No. 2 filter paper. The obtained filtrate was brought into contact with 30 g of activated carbon (Kuraray Coal GLC, manufactured by Kuraray Chemical Co., Ltd.), followed by filtration through a 0.2 μm membrane filter. Finally, 200 g of ion-exchanged water was added, ethanol was distilled off at 40 ° C. and 2.7 kPa, and then the water content was adjusted to obtain “purified green tea extract”. The obtained extract contained 20.2% by mass of non-polymer catechin, and the recovery rate of non-polymer catechins from the tannase-treated green tea extract of Example 2 was 60.5%. The gallate body ratio of the catechin composition was 29.3 mass%. The amount of caffeine was 0.73% by mass and the amount of gallic acid was 2.56% by mass. It was 56.6% by mass of non-polymer catechins in the solid content of the tea extract.
実施例１〜５、比較例１及び２記載の茶抽出物を食品衛生法に基づく殺菌処理をし、風味及び苦味を評価した。その結果を表２に示す。 The tea extracts described in Examples 1 to 5 and Comparative Examples 1 and 2 were sterilized based on the Food Sanitation Law and evaluated for flavor and bitterness. The results are shown in Table 2.
実施例１〜５では処理前後における非重合体カテキン類の回収率が高く、非重合体カテキンガレート率が減少し、没食子酸を殆ど含まず、カフェイン濃度が低減した茶抽出物を得ることができた。更に実施例６では、外観、呈味の安定性が良い飲料が製造できた。比較例１では酸味と雑味が、比較例２では苦味が残る。比較例３ではカフェイン含量が多く、比較例４では酸味が残る。 In Examples 1 to 5, the recovery rate of non-polymer catechins before and after the treatment is high, the non-polymer catechin gallate rate is reduced, and a tea extract containing almost no gallic acid and having a reduced caffeine concentration can be obtained. did it. Furthermore, in Example 6, a beverage having good appearance and stable taste could be produced. In Comparative Example 1, sourness and miscellaneous taste remain, and in Comparative Example 2, bitterness remains. Comparative Example 3 has a high caffeine content, and Comparative Example 4 has a sour taste.
実施例６
実施例３の精製緑茶抽出物を用いて表３に記載の容器詰飲料を調製した。食品衛生法に基づく殺菌処理及びホットパック充填を行って容器詰飲料とした。
Example 6
Using the purified green tea extract of Example 3, the packaged beverages shown in Table 3 were prepared. A sterilization treatment and hot pack filling based on the Food Sanitation Law were performed to obtain a container-packed beverage.
製造した容器詰飲料を３７℃で３０日間保存した後、評価した。外観、呈味の安定性は良かった。 The produced packaged beverage was stored at 37 ° C. for 30 days and then evaluated. Appearance and taste stability were good.
Claims (4)
（ａ）固形分中の非重合体カテキン類２５〜９５質量％
（ｂ）非重合体カテキンガレート体０〜７０質量％
（ｃ）没食子酸と非重合体カテキン類との比率が０〜０．１
（ｄ）カフェインと非重合体カテキン類の比率が０〜０．１５ After the tea extract is hydrolyzed , it is adsorbed on a synthetic adsorbent, and the synthetic adsorbent is washed with an organic solvent aqueous solution having an organic solvent concentration of 0 to 20% by mass. A method for producing a purified tea extract having the following (a) to (d), wherein catechins are eluted.
(A ) Non-polymer catechins in solid content of 25 to 95% by mass
( B) Non-polymer catechin gallate body 0-70 mass%
( C) The ratio of gallic acid to non-polymer catechins is 0 to 0.1
( D) The ratio of caffeine to non-polymer catechins is 0 to 0.15
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| KR1020087021446A KR101436645B1 (en) | 2006-03-02 | 2007-03-01 | Process for producing purified tea extract |
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