JPH0656884A - Purification of glutathione - Google Patents
Purification of glutathioneInfo
- Publication number
- JPH0656884A JPH0656884A JP23262792A JP23262792A JPH0656884A JP H0656884 A JPH0656884 A JP H0656884A JP 23262792 A JP23262792 A JP 23262792A JP 23262792 A JP23262792 A JP 23262792A JP H0656884 A JPH0656884 A JP H0656884A
- Authority
- JP
- Japan
- Prior art keywords
- glutathione
- acetic acid
- exchange resin
- weakly basic
- basic anion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 title claims abstract description 100
- 108010024636 Glutathione Proteins 0.000 title claims abstract description 50
- 229960003180 glutathione Drugs 0.000 title claims abstract description 50
- 238000000746 purification Methods 0.000 title description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 78
- 239000003957 anion exchange resin Substances 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims description 20
- 238000005342 ion exchange Methods 0.000 claims description 4
- 229920000768 polyamine Polymers 0.000 claims description 4
- 239000003480 eluent Substances 0.000 claims description 2
- PABVKUJVLNMOJP-WHFBIAKZSA-N Glu-Cys Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CS)C(O)=O PABVKUJVLNMOJP-WHFBIAKZSA-N 0.000 abstract description 3
- 239000012535 impurity Substances 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 13
- 238000004128 high performance liquid chromatography Methods 0.000 description 8
- 125000000218 acetic acid group Chemical class C(C)(=O)* 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 239000007788 liquid Substances 0.000 description 4
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 2
- 229930189936 Glyoxalase Natural products 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- RITKHVBHSGLULN-WHFBIAKZSA-N L-gamma-glutamyl-L-cysteine Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(O)=O RITKHVBHSGLULN-WHFBIAKZSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 150000001879 copper Chemical class 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- 108010068906 gamma-glutamylcysteine Proteins 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- -1 In addition Chemical compound 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 241000235646 Cyberlindnera jadinii Species 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- BDAGIHXWWSANSR-UHFFFAOYSA-N Formic acid Chemical group OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical group Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- BERDEBHAJNAUOM-UHFFFAOYSA-N copper(I) oxide Inorganic materials [Cu]O[Cu] BERDEBHAJNAUOM-UHFFFAOYSA-N 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- KRFJLUBVMFXRPN-UHFFFAOYSA-N cuprous oxide Chemical compound [O-2].[Cu+].[Cu+] KRFJLUBVMFXRPN-UHFFFAOYSA-N 0.000 description 1
- 229940112669 cuprous oxide Drugs 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- REFMEZARFCPESH-UHFFFAOYSA-M sodium;heptane-1-sulfonate Chemical compound [Na+].CCCCCCCS([O-])(=O)=O REFMEZARFCPESH-UHFFFAOYSA-M 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 150000003512 tertiary amines Chemical class 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明はグルタチオンの精製法に
関するものである。一般にグルタチオンは酵母及び動物
の肝臓などに広く分布しており、生体内の酸化還元系に
関与しているトリペプタイドで、肝機能回復作用や解毒
作用などの重要な役割を果たす医薬上極めて有用な物質
である。FIELD OF THE INVENTION The present invention relates to a method for purifying glutathione. In general, glutathione is widely distributed in yeast and animal liver, and is a tripeptide involved in the redox system in vivo, and is extremely useful as a drug that plays an important role in liver function recovery and detoxification. It is a substance.
【０００２】[0002]
【従来の技術】従来、グルタチオンの精製法としては以
下の方法が知られている。 １）硫酸酸性下亜酸化銅と銅塩を形成させる方法。 ２）強酸性陽イオン交換樹脂に吸着させ、酸または塩に
より溶離する方法（特公昭４４−２３９号、特公昭４５
−４７５５号、特公昭４６−２８３８号）。 ３）弱塩基性陰イオン交換樹脂を通過させる方法（特公
昭４５−２７７９７号） ４）スチレン−ジビニルベンゼン共重合体よりなる多孔
性非極性樹脂を使用する方法（特開昭４９−１２６８８
９号、特開昭５２−１００４２１号）。 ５）弱塩基性陰イオン交換樹脂に吸着後、酢酸で溶離す
る方法（特開昭６１−２８２３９７号）。2. Description of the Related Art Conventionally, the following methods have been known as methods for purifying glutathione. 1) A method of forming a copper salt with cuprous oxide under sulfuric acid acidity. 2) A method of adsorbing to a strongly acidic cation exchange resin and eluting with an acid or salt (Japanese Patent Publication Nos. 44-239 and 45-45).
-4755, Japanese Patent Publication No. 46-2838). 3) Method of passing a weakly basic anion exchange resin (Japanese Patent Publication No. 27797/45) 4) Method of using a porous non-polar resin composed of a styrene-divinylbenzene copolymer (JP-A-49-12688)
No. 9, JP-A No. 52-100421). 5) A method of adsorbing on a weakly basic anion exchange resin and then eluting with acetic acid (JP-A-61-282397).
【０００３】[0003]
【発明が解決しようとする課題】しかしながら、従来提
案された上記１）〜４）の方法によるグルタチオンの精
製は、不純物であるシステインやγ−グルタミルシステ
インを除去できないという欠点があり、一方、５）の方
法は精製は十分に行えるものの、溶離液として酢酸水溶
液を使用するため、結晶化時に酢酸が大量に含有される
という欠点があった。However, the purification of glutathione by the previously proposed methods 1) to 4) has a drawback in that impurities such as cysteine and γ-glutamylcysteine cannot be removed, while 5). Although the method described above can sufficiently perform purification, it has a drawback that a large amount of acetic acid is contained during crystallization because an acetic acid aqueous solution is used as an eluent.
【０００４】[0004]
【課題を解決するための手段】本発明者らは酢酸を含有
しない高純度のグルタチオンを工業的に製造する方法に
おいて鋭意研究を行った結果、ポリアミンをイオン交換
基とする弱塩基性陰イオン交換樹脂にグルタチオンを含
む液を通すことにより、酢酸を含有しない高純度のグル
タチオンを取得することが可能であることを見出し、本
発明を完成するに至ったものである。即ち、本発明は酢
酸を含有しないか、もしくは酢酸含量が極めて低い、高
純度のグルタチオンの精製法を提供するものである。Means for Solving the Problems The inventors of the present invention have earnestly studied in a method for industrially producing high-purity glutathione containing no acetic acid, and as a result, weakly basic anion exchange using polyamine as an ion exchange group. It was found that high-purity glutathione containing no acetic acid can be obtained by passing a solution containing glutathione through a resin, and the present invention has been completed. That is, the present invention provides a high-purity method for purifying glutathione that does not contain acetic acid or has extremely low acetic acid content.
【０００５】以下本発明について詳しく述べる。本発明
に使用されるグルタチオン含有液は、酵母などの微生物
よりの抽出液、サルベージ合成により得られる反応液ま
たはそれらの部分精製液等生化学的反応により得られる
グルタチオン含有液であればいずれにも用いられる。本
発明に用いられる一段目の弱塩基性陰イオン交換樹脂
は、イオン交換基が３級アミンを有するものが好まし
く、更に、弱塩基性陰イオン交換樹脂のイオン形は酢酸
形、ギ酸形、遊離形またはそれらの混合形が用いられ
る。中でも酢酸形または酢酸形と遊離形との混合形が特
に好ましい。硫酸形及び塩酸形の場合はグルタチオンを
殆ど吸着せず好ましくない。酢酸を除くための二段目の
弱塩基性陰イオン交換樹脂は、イオン交換基がポリアミ
ンであり、そのイオン形は遊離形である。使用されるイ
オン交換樹脂の例としては、ダウエックスＷＧＲ−２
（ダウケミカル社製）、アンバーライトＩＲＡ−６０Ｅ
（ロームアンドハース社製）等を挙げることが出来る。The present invention will be described in detail below. The glutathione-containing solution used in the present invention is an extract from a microorganism such as yeast, a reaction solution obtained by salvage synthesis or a glutathione-containing solution obtained by a biochemical reaction such as a partially purified solution thereof. Used. The first-stage weakly basic anion exchange resin used in the present invention preferably has an ion exchange group having a tertiary amine, and the weakly basic anion exchange resin has an ionic form of an acetic acid form, a formic acid form, a free form. Forms or mixed forms thereof are used. Among them, acetic acid form or a mixed form of acetic acid form and free form is particularly preferable. The sulfuric acid form and the hydrochloric acid form are not preferable because glutathione is hardly adsorbed. The second-stage weakly basic anion exchange resin for removing acetic acid has a polyamine as an ion exchange group and its ionic form is a free form. As an example of the ion exchange resin used, Dowex WGR-2
(Manufactured by Dow Chemical Co.), Amberlite IRA-60E
(Made by Rohm and Haas) and the like.
【０００６】上記グルタチオン含有液を酢酸形または酢
酸形と遊離形の弱塩基性陰イオン交換樹脂を詰めたカラ
ムに、通液速度（ＳＶ）０．５〜３で通液してグルタチ
オンを吸着させる。適当量の水で洗浄した後、酢酸溶
液、続いて水を通液してグルタチオンを溶出させる。こ
の時一段目の弱塩基性陰イオン交換樹脂カラムの下端よ
りグルタチオンの溶出が認められると同時に、溶出液を
二段目のポリアミンを交換基とする陰イオン交換樹脂を
詰めたカラムに導く。二段目のカラムよりの溶出液中の
グルタチオン及び酢酸の濃度推移を、高速液体クロマト
グラフィー（ＨＰＬＣ）またはその他の適当な方法で測
定する事によって、最初にグルタチオンが溶出し、続い
て酢酸が溶出する事がわかる。溶出液の適当な分画部を
集めることによって、酢酸を全く含まないか、もしくは
酢酸濃度が非常に低いグルタチオン溶液を得ることがで
きる。得られたグルタチオン溶液を濃縮し、次いで結晶
化や凍結乾燥することにより高純度のグルタチオンを製
造することができる。The above glutathione-containing liquid is passed through a column packed with an acetic acid form or an acetic acid form and a free form of a weakly basic anion exchange resin at a liquid flow rate (SV) of 0.5 to 3 to adsorb glutathione. . After washing with an appropriate amount of water, glutathione is eluted by passing an acetic acid solution and then water. At this time, glutathione was eluted from the lower end of the first-step weakly basic anion-exchange resin column, and at the same time, the eluate was introduced into a column packed with an anion-exchange resin having a second-stage polyamine as an exchange group. Glutathione and acetic acid are eluted first by measuring the transition of the concentration of glutathione and acetic acid in the eluate from the second column by high performance liquid chromatography (HPLC) or other appropriate method. I know what to do. By collecting appropriate fractions of the eluate, a glutathione solution containing no acetic acid or a very low acetic acid concentration can be obtained. High-purity glutathione can be produced by concentrating the obtained glutathione solution and then crystallizing or freeze-drying.
【０００７】[0007]
【実施例】次に実施例により具体的に本発明を説明する
が、これによって本発明が制限されるものではない。な
お、本実施例中グルタチオンの定量法はヨード法及びグ
リオキサラーゼ法（「メソッド・イン・エンザイモロジ
ー」第１巻５４０頁、アカデミックプレス社、１９５５
年版）で行った。またＨＰＬＣの測定条件は以下の通り
である。 １．カラム：ＴＳＫ ｇｅｌ ＯＤＳ−８０−ＴＭ、長
さ：１５ｃｍ、内径：４．６ｍｍ ２．緩衝液：０．６８６％リン酸二水素カリウム及び
０．０２５％１−ヘプタンスルホン酸ナトリウムを含む
３％メタノール水溶液、ＰＨ３．０ ３．流 速：１．０ｍｌ／ｍｉｎ． ４．検出器：ＵＶ検出器（波長２１０ｎｍ）EXAMPLES The present invention will now be specifically described with reference to examples, but the present invention is not limited thereto. In this Example, glutathione was quantified by iodine method and glyoxalase method (“Method in Enzymology”, Vol. 1, p. 540, Academic Press Co., 1955).
Year edition). The HPLC measurement conditions are as follows. 1. Column: TSK gel ODS-80-TM, length: 15 cm, inner diameter: 4.6 mm 2. Buffer: 3% aqueous methanol solution containing 0.686% potassium dihydrogen phosphate and 0.025% sodium 1-heptanesulfonate, pH 3.0. Flow rate: 1.0 ml / min. 4. Detector: UV detector (wavelength 210nm)
【０００８】実施例１ キャンディダ・ウチルス ＫＪＳ−０５８２株（ＦＥＲ
Ｍ Ｐ−７３９６株）の培養菌体７００ｇ（乾燥時換
算）を熱水抽出し、除菌後常法により銅塩を形成させ、
硫化水素で脱銅することにより、グルタチオン３０．５
ｇ（グリオキサラーゼ法）を含む溶液５００ｍｌを得
た。該グルタチオン含有液を酢酸形とした弱塩基性陰イ
オン交換樹脂ダイヤイオンＷＡ３０（三菱化成工業製）
を詰めたカラム（内径４５ｍｍ、高さ１８０ｍｍ）に、
ＳＶ＝１で通液し、５００ｍｌの水で洗浄した。次いで
６．０％の酢酸水溶液２６０ｍｌを用いＳＶ＝１で溶離
した。さらに水で溶離を続けた。カラムよりグルタチオ
ンの溶出が始まると同時に、直列に接続した弱塩基性陰
イオン交換樹脂ダウエックスＷＧＲ−２（ダウケミカル
社製）を詰めたカラム（内径３０ｍｍ、高さ１７０ｍ
ｍ）に通液する。本カラムクロマトグラフィーにおける
グルタチオン及び酢酸の溶離の様子を図１に示す。得ら
れたグルタチオン画分を減圧濃縮することにより結晶グ
ルタチオン２２．０ｇを得た。得られた結晶グルタチオ
ンをＨＰＬＣで分析した結果、γ−グルタミルシステイ
ンや酢酸を殆ど含んでいなかった。その測定結果を表１
に示す。また、得られた結晶グルタチオンのＨＰＬＣチ
ャートを図２に示す。 比較例 実施例１と同様にして得られたカラム処理前の液を、酢
酸形とした弱塩基性陰イオン交換樹脂ダイヤイオンＷＡ
３０を詰めたカラム（内径４５ｍｍ、高さ１８０ｍｍ）
に、ＳＶ＝１で通液し、５００ｍｌの水で洗浄した。次
いで６．０％の酢酸水溶液２６０ｍｌ更に水を用いＳＶ
＝１で溶離しグルタチオン画分４００ｍｌを得た。グル
タチオンを含む画分をそのまま減圧濃縮し、結晶化する
ことにより、結晶グルタチオン２４．４ｇを得た。得ら
れた結晶グルタチオンをＨＰＬＣで分析した結果、γ−
グルタミルシステイン及びシステインは含まないものの
多量の酢酸の混入が認められた。この様にして得られた
結晶グルタチオン及び実施例１で得られた結晶グルタチ
オンについて、ＨＰＬＣで測定したグルタチオンの純度
及びグルタチオン中に含まれる酢酸の濃度を表１に示
す。Example 1 Candida utilis KJS-0582 strain (FER
700 g of cultured bacterial cells (MP-7396 strain) (equivalent to dry time) were extracted with hot water to form a copper salt by a conventional method after sterilization.
By decopperizing with hydrogen sulfide, glutathione 30.5
500 ml of a solution containing g (glyoxalase method) was obtained. Weakly basic anion exchange resin Diaion WA30 (manufactured by Mitsubishi Kasei Co., Ltd.) in which the glutathione-containing solution is in the acetic acid form
In a column packed with (inner diameter 45 mm, height 180 mm)
The solution was passed at SV = 1 and washed with 500 ml of water. Then, it was eluted with SV = 1 using 260 ml of a 6.0% acetic acid aqueous solution. Further elution was continued with water. At the same time that glutathione begins to elute from the column, a column (inner diameter 30 mm, height 170 m) packed with a series of weakly basic anion exchange resin Dowex WGR-2 (manufactured by Dow Chemical Co.)
Pour into m). The state of elution of glutathione and acetic acid in this column chromatography is shown in FIG. The obtained glutathione fraction was concentrated under reduced pressure to obtain 22.0 g of crystalline glutathione. The obtained crystalline glutathione was analyzed by HPLC and as a result, it contained almost no γ-glutamylcysteine or acetic acid. The measurement results are shown in Table 1.
Shown in. The HPLC chart of the obtained crystalline glutathione is shown in FIG. Comparative Example The liquid before column treatment obtained in the same manner as in Example 1 was converted to an acetic acid form, a weakly basic anion exchange resin DIAION WA.
Column packed with 30 (inner diameter 45 mm, height 180 mm)
The solution was passed through SV = 1 and washed with 500 ml of water. Then, 260 ml of a 6.0% acetic acid aqueous solution and SV using water.
Elution with = 1 gave 400 ml of glutathione fraction. The fraction containing glutathione was directly concentrated under reduced pressure and crystallized to obtain 24.4 g of crystalline glutathione. As a result of analyzing the obtained crystalline glutathione by HPLC, γ-
Although glutamyl cysteine and cysteine were not included, a large amount of acetic acid was found to be mixed. With respect to the crystalline glutathione thus obtained and the crystalline glutathione obtained in Example 1, the purity of glutathione measured by HPLC and the concentration of acetic acid contained in glutathione are shown in Table 1.
【０００９】[0009]
【表１】 ──────────────────────────────────── 試 料 ＨＰＬＣ純度 酢酸濃度 （％） （ＰＰＭ） ──────────────────────────────────── 本発明による結晶グルタチオン ９８．５ １０ ──────────────────────────────────── 比較例で得られた結晶グルタチオン ９８．１ ４５０ ────────────────────────────────────[Table 1] ──────────────────────────────────── Samples HPLC purity Acetic acid concentration (%) ( PPM) ──────────────────────────────────── Crystalline glutathione according to the present invention 98.5 10 ─── ────────────────────────────────── Crystalline glutathione obtained in Comparative Example 98.1 450 ───── ───────────────────────────────
【００１０】[0010]
【図１】[Figure 1]
【００１１】[0011]
【図２】[Fig. 2]
【００１２】[0012]
【発明の効果】以上説明してきたように、本発明の精製
法によればグルタミルシステイン等の不純物を含まず、
且つ酢酸をほとんど含有しないグルタチオンを工業的に
有利に取得することが出来る。As described above, the purification method of the present invention does not contain impurities such as glutamylcysteine,
In addition, glutathione containing almost no acetic acid can be obtained industrially advantageously.
【図１】弱塩基性陰イオン交換樹脂ダウエックスＷＧＲ
−２によるグルタチオン含有液の分画の様子を示したも
のである。Figure 1: Weakly basic anion exchange resin Dowex WGR
2 shows a state of fractionation of a glutathione-containing liquid by -2.
【図２】弱塩基性陰イオン交換樹脂ダウエックスＷＧＲ
−２により分画したグルタチオン画分を減圧濃縮するこ
とにより得られた結晶グルタチオンのＨＰＬＣ測定チャ
ートを示したものである。Fig. 2 Weakly basic anion exchange resin Dowex WGR
2 shows an HPLC measurement chart of crystalline glutathione obtained by concentrating the glutathione fraction fractionated by -2 under reduced pressure.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 松井 勝明 大分県佐伯市大字池田1411−４ (72)発明者 日野 忠行 東京都三鷹市下連雀６−14−６−23 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Katsuaki Matsui 1411-4 Ikeda, Saiki City, Oita Prefecture (72) Inventor Tadayuki Hino 6-14-6-23 Shimorenjaku, Mitaka City, Tokyo
Claims (2)
ン交換樹脂に通した後、酢酸水溶液でグルタチオンを溶
離し、次いで溶離液を別の弱塩基性陰イオン交換樹脂に
通液することにより酢酸を除くことを特徴とするグルタ
チオンの精製法。1. A solution containing glutathione is passed through a weakly basic anion exchange resin, glutathione is eluted with an aqueous acetic acid solution, and then the eluent is passed through another weakly basic anion exchange resin to obtain acetic acid. A method for purifying glutathione, characterized in that
樹脂のイオン交換基がポリアミンである請求項１項記載
のグルタチオンの精製法。2. The method for purifying glutathione according to claim 1, wherein the ion exchange group of the weakly basic anion exchange resin for removing acetic acid is polyamine.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23262792A JP3315158B2 (en) | 1992-08-10 | 1992-08-10 | Glutathione purification method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23262792A JP3315158B2 (en) | 1992-08-10 | 1992-08-10 | Glutathione purification method |
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| Publication Number | Publication Date |
|---|---|
| JPH0656884A true JPH0656884A (en) | 1994-03-01 |
| JP3315158B2 JP3315158B2 (en) | 2002-08-19 |
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ID=16942288
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP23262792A Expired - Fee Related JP3315158B2 (en) | 1992-08-10 | 1992-08-10 | Glutathione purification method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003035674A1 (en) * | 2001-10-25 | 2003-05-01 | Kyowa Hakko Kogyo Co., Ltd. | Crystal of oxidized glutathione and process for producing the same |
| WO2008001837A1 (en) * | 2006-06-28 | 2008-01-03 | Kyowa Hakko Bio Co., Ltd. | Method for purification of oligopeptide |
| CN100455592C (en) * | 2006-05-19 | 2009-01-28 | 江南大学 | A method for extracting glutathione from glutathione fermentation broth |
| WO2011132724A1 (en) * | 2010-04-21 | 2011-10-27 | 協和発酵バイオ株式会社 | Crystalline oxidized glutathione and production method therefor |
| CN113853114A (en) * | 2019-03-29 | 2021-12-28 | 株式会社钟化 | Composition comprising glutathione for application to leaves of plants |
| CN114560907A (en) * | 2022-02-23 | 2022-05-31 | 武汉绿合医药科技有限公司 | Preparation method of glutathione impurity |
-
1992
- 1992-08-10 JP JP23262792A patent/JP3315158B2/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003035674A1 (en) * | 2001-10-25 | 2003-05-01 | Kyowa Hakko Kogyo Co., Ltd. | Crystal of oxidized glutathione and process for producing the same |
| US7094870B2 (en) | 2001-10-25 | 2006-08-22 | Kyowa Hakko Kogyo Co., Ltd. | Crystals of oxidized glutathione and process for producing the same |
| CN100455592C (en) * | 2006-05-19 | 2009-01-28 | 江南大学 | A method for extracting glutathione from glutathione fermentation broth |
| WO2008001837A1 (en) * | 2006-06-28 | 2008-01-03 | Kyowa Hakko Bio Co., Ltd. | Method for purification of oligopeptide |
| WO2011132724A1 (en) * | 2010-04-21 | 2011-10-27 | 協和発酵バイオ株式会社 | Crystalline oxidized glutathione and production method therefor |
| US9000126B2 (en) | 2010-04-21 | 2015-04-07 | Kyowa Hakko Bio Co., Ltd. | Crystalline oxidized glutathione and production method therefor |
| US9243029B2 (en) | 2010-04-21 | 2016-01-26 | Kyowa Hakko Bio Co., Ltd. | Crystalline oxidized glutathione and production method therefor |
| CN113853114A (en) * | 2019-03-29 | 2021-12-28 | 株式会社钟化 | Composition comprising glutathione for application to leaves of plants |
| CN113853114B (en) * | 2019-03-29 | 2024-06-11 | 株式会社钟化 | Composition comprising glutathione for application to leaves of plants |
| CN114560907A (en) * | 2022-02-23 | 2022-05-31 | 武汉绿合医药科技有限公司 | Preparation method of glutathione impurity |
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| Publication number | Publication date |
|---|---|
| JP3315158B2 (en) | 2002-08-19 |
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