JPH01213258A - Purification of carnitine and carnitine amide - Google Patents
Purification of carnitine and carnitine amideInfo
- Publication number
- JPH01213258A JPH01213258A JP63038690A JP3869088A JPH01213258A JP H01213258 A JPH01213258 A JP H01213258A JP 63038690 A JP63038690 A JP 63038690A JP 3869088 A JP3869088 A JP 3869088A JP H01213258 A JPH01213258 A JP H01213258A
- Authority
- JP
- Japan
- Prior art keywords
- carnitine
- carnitinamide
- resin
- aqueous solution
- amide
- 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.)
- Pending
Links
- 229960004203 carnitine Drugs 0.000 title claims abstract description 51
- KWIXGIMKELMNGH-UHFFFAOYSA-O carnitinamide Chemical compound C[N+](C)(C)CC(O)CC(N)=O KWIXGIMKELMNGH-UHFFFAOYSA-O 0.000 title claims abstract description 49
- 238000000746 purification Methods 0.000 title claims abstract description 5
- PHIQHXFUZVPYII-ZCFIWIBFSA-O (R)-carnitinium Chemical compound C[N+](C)(C)C[C@H](O)CC(O)=O PHIQHXFUZVPYII-ZCFIWIBFSA-O 0.000 title 1
- PHIQHXFUZVPYII-ZCFIWIBFSA-N (R)-carnitine Chemical compound C[N+](C)(C)C[C@H](O)CC([O-])=O PHIQHXFUZVPYII-ZCFIWIBFSA-N 0.000 claims abstract description 60
- 239000011347 resin Substances 0.000 claims abstract description 34
- 229920005989 resin Polymers 0.000 claims abstract description 34
- 239000007864 aqueous solution Substances 0.000 claims abstract description 27
- 239000003729 cation exchange resin Substances 0.000 claims abstract description 17
- 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 claims abstract description 15
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims abstract description 8
- 150000001412 amines Chemical class 0.000 claims abstract description 8
- 150000003863 ammonium salts Chemical class 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 13
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- -1 amine salt Chemical class 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims 1
- 239000000243 solution Substances 0.000 abstract description 10
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 abstract description 9
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract description 2
- 235000014113 dietary fatty acids Nutrition 0.000 abstract description 2
- 239000000194 fatty acid Substances 0.000 abstract description 2
- 229930195729 fatty acid Natural products 0.000 abstract description 2
- 150000004665 fatty acids Chemical class 0.000 abstract description 2
- 210000000748 cardiovascular system Anatomy 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 238000002955 isolation Methods 0.000 abstract 1
- 239000012528 membrane Substances 0.000 abstract 1
- 210000003470 mitochondria Anatomy 0.000 abstract 1
- 229940124597 therapeutic agent Drugs 0.000 abstract 1
- 238000011084 recovery Methods 0.000 description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 230000002378 acidificating effect Effects 0.000 description 6
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 5
- 239000005695 Ammonium acetate Substances 0.000 description 5
- 235000019257 ammonium acetate Nutrition 0.000 description 5
- 229940043376 ammonium acetate Drugs 0.000 description 5
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 5
- 238000005406 washing Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- ALSPKRWQCLSJLV-UHFFFAOYSA-N azanium;acetic acid;acetate Chemical compound [NH4+].CC(O)=O.CC([O-])=O ALSPKRWQCLSJLV-UHFFFAOYSA-N 0.000 description 3
- 238000011109 contamination Methods 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- AWADHHRPTLLUKK-UHFFFAOYSA-N diazanium sulfuric acid sulfate Chemical compound [NH4+].[NH4+].OS(O)(=O)=O.[O-]S([O-])(=O)=O AWADHHRPTLLUKK-UHFFFAOYSA-N 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- QUSNBJAOOMFDIB-UHFFFAOYSA-N Ethylamine Chemical compound CCN QUSNBJAOOMFDIB-UHFFFAOYSA-N 0.000 description 2
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 239000002948 appetite stimulant Substances 0.000 description 2
- 229940023913 cation exchange resins Drugs 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 2
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 239000000052 vinegar Substances 0.000 description 2
- PHIQHXFUZVPYII-LURJTMIESA-N (S)-carnitine Chemical compound C[N+](C)(C)C[C@@H](O)CC([O-])=O PHIQHXFUZVPYII-LURJTMIESA-N 0.000 description 1
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102100022653 Histone H1.5 Human genes 0.000 description 1
- 101000899879 Homo sapiens Histone H1.5 Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- DKAGJZJALZXOOV-UHFFFAOYSA-N hydrate;hydrochloride Chemical compound O.Cl DKAGJZJALZXOOV-UHFFFAOYSA-N 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000001700 mitochondrial membrane Anatomy 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 235000011056 potassium acetate Nutrition 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
(イ)産業上の利用分野
本発明はカルニチンアミドを含有するカルニチン水溶液
から陽イオン交換樹脂を用いて両者を分離精製する方法
に関する。DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application The present invention relates to a method for separating and purifying carnitinamide from an aqueous carnitine solution using a cation exchange resin.
カルニチン(−一ヒドロキシーγ−トリメチルアミノ酪
fi)にtiL体およびD体の光学異性体が存在する。Carnitine (-monohydroxy-γ-trimethylaminobutyrofi) exists as tiL and D optical isomers.
−一カルニチンは通常生体内に存在し脂肪酸のミトコン
ドリア膜通過の際のキャリアーとして作用していること
が明らかとなっている。-It has been revealed that monocarnitine normally exists in living organisms and acts as a carrier when fatty acids pass through the mitochondrial membrane.
カルニチン類の治療学士の用途としては従来食欲促進剤
などにラセミ体が用いられてきたが、近年、心血1!P
系での病気の分野や腎臓系の病気の分野において5体の
みの使用が効果的であることが判明し、′L体に対する
関心が高まってきている。Conventionally, the racemic form of carnitine has been used as an appetite stimulant, but in recent years, it has been used as an appetite stimulant. P
It has been found that the use of only 5 forms is effective in the fields of diseases related to the kidney system and diseases of the kidney system, and interest in the 'L form is increasing.
←)従来の技術
り一カルニチンを製造°する方法は数多く知られている
か、その中でDL−カルニチンアミドを微生物の生産す
る酵素で不斉加水分解させてL−カルニチンを取得する
方法(特願昭6l−198173)においては得られる
L−カルニチン水浴液中に残存するD−カルニチンアミ
ドを完全に除去する必要がある。これまでカルニチンと
カルニチンアミドの混合水溶液から両者を分離、精製す
る方法は知られていない。←) Conventional technology: There are many known methods for producing carnitine. Among them, there is a method in which L-carnitine is obtained by asymmetrically hydrolyzing DL-carnitinamide with an enzyme produced by microorganisms (patent application). In Sho 6l-198173), it is necessary to completely remove D-carnitinamide remaining in the L-carnitine water bath solution obtained. Until now, there is no known method for separating and purifying carnitine and carnitinamide from a mixed aqueous solution.
(ハ)発明が解決しようとする問題点と問題点を解決す
るだめの手段
これまで知られているカルニチンの精製法の代表例とし
てはH+形の陽イオン交換樹脂にカルニチンを吸着した
後、アンモニア水や稀塩酸で沈埋してカルニチンを溶離
、精製する方法がある。しかし、H+形の陽イオン交換
樹脂にカルニチンアミドを接触させるとカルニチンアミ
ドの大半がカルニチンへ分解するので、この方法を用い
てL−カルニチンとD−カルニチンアミドの混合水溶液
から両者を分離できない。−方、H+形以外のイオン形
の陽イオン交換樹脂、聞えは、Na+形、X+形やMu
−形などの陽イオン交換樹脂にはカルニチンアミドは吸
着されるがカルニチンはほとんど吸着されない。このこ
とを利用してカルニチンとカルニチンアミドの混合水溶
液から両者を分離、#I製することは可能ではあるが、
非吸着のカルニチンの精製工程を設ける必要があるなど
工業的には満足いくものではない。(c) Problems to be solved by the invention and means to solve the problems A representative example of the known purification method for carnitine is to adsorb carnitine on an H+ type cation exchange resin, and then use ammonia There is a method to elute and purify carnitine by embedding it in water or dilute hydrochloric acid. However, when carnitinamide is brought into contact with a cation exchange resin in the H+ form, most of the carnitinamide decomposes into carnitine, so this method cannot be used to separate L-carnitine and D-carnitinamide from a mixed aqueous solution. Cation exchange resins in ionic form other than H+ form, Na+ form, X+ form, and Mu
Carnitinamide is adsorbed on cation exchange resins such as - type, but carnitine is hardly adsorbed. Although it is possible to utilize this fact to separate carnitine and carnitinamide from a mixed aqueous solution and produce #I,
This method is not industrially satisfactory, as it requires a purification process for non-adsorbed carnitine.
さらに、樹脂に吸着したカルニチンアミドは濃アンモニ
ア水で処理しても溶離できず、また、稀塩酸や稀硫酸な
どで処理するとそれらの濃度によっては溶離時にカルニ
チンアミドがカルニチンに分解してしまうなどカル−チ
ンアミドの精製上にも難点があった。Furthermore, carnitinamide adsorbed to the resin cannot be eluted even when treated with concentrated ammonia water, and when treated with dilute hydrochloric acid or dilute sulfuric acid, carnitinamide decomposes into carnitine during elution depending on their concentration. - There were also difficulties in purifying tinamide.
本発明者らはカルニチンとカルニチンアミドを効率良く
分離、精製する方法について鋭意研究を行った結果、p
H1〜5の塩類の水溶液で平衡化させた陽イオン交換樹
脂を用いることによって両者を効率曳く分離、精製する
方法を見い出した。The present inventors conducted intensive research on a method for efficiently separating and purifying carnitine and carnitinamide, and as a result, p
By using a cation exchange resin equilibrated with an aqueous solution of salts of H1-5, we have found a method for efficiently separating and purifying both.
に)作用
本発明によれば、カルニチンアミドはカルニチンへのシ
無分解を9けることなく陽イオン交換樹脂に吸着し、ま
た、カルニチンへ分解されることなく溶離される丸め、
L−カルニチンとD−カルニチンアミドの混合水溶液か
ら両者を分離する際、D−カルニチンアミドの分解によ
り生成するD−カルニチンの混入によるL−カルニチン
の光学純度の低下を防ぐことが可能となる。According to the present invention, carnitinamide is adsorbed on a cation exchange resin without being decomposed into carnitine, and is eluted without being decomposed into carnitine.
When separating L-carnitine and D-carnitinamide from a mixed aqueous solution, it is possible to prevent a decrease in the optical purity of L-carnitine due to contamination with D-carnitine produced by decomposition of D-carnitinamide.
本発明において用いる陽イオン交換樹脂は通常の予備処
理まfc、は再生処理後、酢酸アンモニウム−酢酸や硫
酸アンモニウム−硫酸などのpH1〜5の水溶液で平衡
化させることができるが、平衡化水溶液のpHiが1〜
50間であればこれらの水溶液以外の燐酸アンモニウム
、塩化アンモニウム、酢酸ナトリウム、燐酸ナトリウム
、酢酸カリウム、燐酸カリウムやその他の有機酸塩およ
び無銭塩の水溶液を用いることによってもカルニチンと
カルニチンアミドをともに樹脂に吸着させることができ
る。The cation exchange resin used in the present invention can be equilibrated with an aqueous solution of pH 1 to 5 such as ammonium acetate-acetic acid or ammonium sulfate-sulfuric acid after normal pretreatment or regeneration treatment, but the pH of the equilibrated aqueous solution is 1~
For a period of 50 minutes, carnitine and carnitinamide can be combined with the resin by using aqueous solutions of ammonium phosphate, ammonium chloride, sodium acetate, sodium phosphate, potassium acetate, potassium phosphate, and other organic acid salts and free salts other than these aqueous solutions. can be adsorbed to.
カルニチンとカルニチンアミドを吸着させた樹脂をアン
モニア水で処理するとカル−チンアミドを該樹脂に残存
させたままカルニチンのみを特異的に溶離させることが
できる。また、各種アンモニウム塩やアミンの水溶液を
用いてもカル−チンを優先的に溶離させることができる
。When a resin adsorbed with carnitine and carnitinamide is treated with aqueous ammonia, only carnitine can be specifically eluted while carnitine remains in the resin. Furthermore, cartin can be preferentially eluted using aqueous solutions of various ammonium salts or amines.
この場合には、カルニチンを効率よく回収するため、カ
ルニチンアミドの溶離を許さない比較的低濃度の水溶液
を用いる。この濃度はアンモニウム塩やアミンの種類に
よって異なるが、通常1M以下の濃度が好適でめる。In this case, in order to efficiently recover carnitine, an aqueous solution with a relatively low concentration that does not allow elution of carnitinamide is used. Although this concentration varies depending on the type of ammonium salt or amine, a concentration of 1M or less is usually suitable.
カルニチンの溶離が終了したのち樹脂に残存しているカ
ルニチンアミドは各種アンモニウム塩またはアミンやア
ミンの塩の水溶液の濃度を上昇させて処理することによ
シカルニチンへの分解をうけることなく樹脂よシはぼ定
量的に溶離回収できる。After the elution of carnitine is completed, the carnitinamide remaining in the resin can be removed from the resin without being decomposed into cycarnitine by increasing the concentration of various ammonium salts or amines or aqueous solutions of amine salts. It can be eluted and recovered quantitatively.
ここでアミンとしてはトリメチルアミン、トリエチルア
ミン、ジメチルアミン、ジエチルアミン、モノメチルア
ミン、モノエチルアミンなどの各種のアミンを用いるこ
とができる。As the amine, various amines such as trimethylamine, triethylamine, dimethylamine, diethylamine, monomethylamine, and monoethylamine can be used.
また、陽イオン交換樹脂としてはゲル型樹脂のほかマク
ロポーラス型樹脂を用いてもカルニチンとカルニチンア
ミドを同様に分離することができる。Further, carnitine and carnitinamide can be similarly separated using a macroporous resin in addition to a gel type resin as the cation exchange resin.
樹脂から溶離させたカルニチン画分を濃縮後、乾燥させ
ることによシカルニチン分子内塩を得ることができる。Cycarnitine inner salt can be obtained by concentrating and drying the carnitine fraction eluted from the resin.
また、カル−チンを塩酸塩等の各種の塩の形として得る
場合には樹脂から溶離させたカルニチン画分からアンモ
ニアが無くなるまで濃縮させたのち等モルの各種の酸を
加え、再濃縮し、エタノール/アセトン混液を加え、冷
却、結晶化させればよい。In addition, when obtaining carnitine in the form of various salts such as hydrochloride, the carnitine fraction eluted from the resin is concentrated until no ammonia is present, then equimolar amounts of various acids are added, reconcentrated, and ethanol is added. / Acetone mixture may be added, cooled, and crystallized.
一方、溶離させたカルニチンアミド画分からのカルニチ
ンアミドの調製法はガえば特公昭3B−25tlC記滅
された方法に準拠して行うことができる。On the other hand, carnitinamide can be prepared from the eluted carnitinamide fraction according to the method described in Japanese Patent Publication No. 3B-25TLC.
次に本発明の実施例を示す。Next, examples of the present invention will be shown.
実施例1
カルニチン塩酸塩1tとカルニチンアミド塩酸塩1fを
10mM酢酸アンモニウム−酢酸(pH五〇)に溶解し
、pHを五〇に補正後、1011M酢酸アンモニウム−
酢酸(pH五〇)で平衡化させた50m(の強酸性陽イ
オン交換耐脂(ダイヤイオンBK IB )を充填した
ガラスカラム(φ12■工、D、)に通過させた。次に
カラムを水洗した後2囁アンモニア水を通液したところ
カルニチンだけが溶離された。(回収率100僑)
さらに樹脂に残存したカルニチンアミドを8憾酢酸アン
モニウム水溶液を通液することによシ溶離回収した。(
回収率80%)
実施f12
カルニチン塩酸塩1fとカルニチンアミド塩酸塩1ft
−350mM硫酸アンモニウム−硫酸(pH2−o)に
溶解し、pBt−2,0に補正後、501EIM硫酸ア
ンモニウムー硫酸(pH2,0)で平衡化させたtjl
i酸性陽イオン交換樹脂(ダイヤイオン811B )
50−を充填したガラスカラム(φ12■1. D、
)に通過させた。次にカラムを水洗した後2係アンモニ
ア水を通液したところカルニチンだけが溶離された。(
回収率100繋)
さらに樹脂に残存したカルニチンアミドを8%酢酸アン
モニウム水溶液を通液することによ)溶離回収した。(
回収率82僑)
実施例3
実施例2と同様に1PH2,0に平衡化させた樹脂を入
れたカラムにカルニチン塩酸塩とカルニチンアミド塩酸
塩をおのおの18ft含む混合液を通液、樹脂を水洗後
、2優アンモニア水を用いてカルニチンのみを特異的に
溶離させた。Example 1 1t of carnitine hydrochloride and 1f of carnitinamide hydrochloride were dissolved in 10mM ammonium acetate-acetic acid (pH 50), and after correcting the pH to 50, 1011M ammonium acetate-acetic acid was dissolved.
It was passed through a glass column (φ12 mm, D) packed with 50 m of strongly acidic cation exchange fat-resistant (Diaion BK IB) equilibrated with acetic acid (pH 50).Then, the column was washed with water. After that, only carnitine was eluted by passing 2 drops of ammonia water through the resin. (Recovery rate: 100) Furthermore, carnitinamide remaining in the resin was eluted and recovered by passing 8 drops of ammonium acetate aqueous solution. (
Recovery rate 80%) Implementation f12 Carnitine hydrochloride 1f and carnitinamide hydrochloride 1ft
-tjl dissolved in 350mM ammonium sulfate-sulfuric acid (pH 2-o), corrected to pBt-2,0, and equilibrated with 501EIM ammonium sulfate-sulfuric acid (pH 2,0)
i Acidic cation exchange resin (Diaion 811B)
A glass column (φ12■1.D,
) was passed. Next, after washing the column with water, aqueous ammonia was passed through the column, and only carnitine was eluted. (
Further, carnitinamide remaining in the resin was eluted and recovered by passing an 8% ammonium acetate aqueous solution through the resin. (
Example 3 A mixture containing 18 ft of each of carnitine hydrochloride and carnitinamide hydrochloride was passed through a column containing a resin equilibrated to 1PH 2.0 in the same manner as in Example 2, and the resin was washed with water. , only carnitine was specifically eluted using aqueous ammonia.
(回収率100%)
次に樹脂を水洗後、5暢硫酸アンモニウム水溶液を通液
することによシカルニチンアミドを樹脂から溶離した。(Recovery rate: 100%) Next, after washing the resin with water, cycarnitinamide was eluted from the resin by passing a 50% ammonium sulfate aqueous solution through the resin.
(回収率10096)この画分に水酸化カルシウムを加
え4℃に放置、生じた沈澱をp別したのちF液を濃縮し
残留物にエタノール水溶液を加え冷却するとカルニチン
アミド硫酸塩の結晶2.5tt−得た。(回収率50%
)
実施列4
実施911と同様にしてカルニチンとカルニチンアミド
の混合水溶液を通液、カラムを水洗後、1%トリメチル
アミン水溶液を通液することによりカル−チンだけを樹
脂から溶離した。(回収率1oo%)
次に樹脂に残存するカルニチンアミドt−3%トリメチ
ルアミン−塩I!i!(pH9,0)を通液することに
よシ溶離した。(回収率93%)実施列5
実施例1と同様なカルニチンとカルニチンアミドの混合
溶液を10mM#、eIRアンモニウムー酢#1.Cp
H五〇)で平衡化させた50−の強酸性陽イオン交換樹
脂(ダイヤイオン8K IB )を充填したガラスカラ
ムに通過、同一水溶液でカラムを洗浄後、(L4%(2
)酢酸アンモニウム水溶液を通液することによタカル二
チンを樹脂から溶離した。(回収率95%)
次に8%酢酸アンモニウム水溶液を通液することによシ
カルニチンアミドを溶離した。(回収率79%)
実施列6
強酸性陽イオン交換樹脂としてダウエックスHOR−W
2を用い、実施例5と同様な操作を行いカルニチンとカ
ルニチンアミドを分離溶出させた。回収率はカルニチン
が95%、カルニチンアミドが74%であった。(Recovery rate 10096) Calcium hydroxide was added to this fraction and left at 4°C. After separating the resulting precipitate, the F solution was concentrated, and an ethanol aqueous solution was added to the residue. When cooled, 2.5 tt of carnitinamide sulfate crystals were obtained. -I got it. (Recovery rate 50%
) Run 4 In the same manner as in Run 911, a mixed aqueous solution of carnitine and carnitinamide was passed through the column, the column was washed with water, and then only cartin was eluted from the resin by passing a 1% aqueous trimethylamine solution through the column. (Recovery rate 1oo%) Next, carnitinamide t-3% trimethylamine salt I remaining in the resin! i! (pH 9,0) was eluted by passing through the solution. (Recovery rate 93%) Example row 5 A mixed solution of carnitine and carnitinamide similar to that in Example 1 was mixed with 10 mM #, eIR ammonium-vinegar #1. Cp
Passed through a glass column packed with a 50-strong acidic cation exchange resin (Diaion 8K IB) equilibrated with H50). After washing the column with the same aqueous solution,
) Tacarnithine was eluted from the resin by passing an aqueous ammonium acetate solution through the resin. (Recovery rate 95%) Next, cycarnitinamide was eluted by passing an 8% aqueous ammonium acetate solution through the solution. (Recovery rate 79%) Practical row 6 DOWEX HOR-W as a strong acidic cation exchange resin
2 and the same operation as in Example 5 was carried out to separate and elute carnitine and carnitinamide. The recovery rate was 95% for carnitine and 74% for carnitinamide.
実施列7
強酸性陽イオン交換樹脂としてマクロポーラス型(ダウ
エックス 88)を用い、実m915と同様な操作でカ
ル−チンとカルニチンアミドを分離溶出させた。回収率
はカルニチンが96優、カルニチンアミドが84%であ
った。Implementation row 7 Using a macroporous type (Dowex 88) as a strongly acidic cation exchange resin, cartin and carnitinamide were separated and eluted in the same manner as in the actual m915. The recovery rate was 96% for carnitine and 84% for carnitinamide.
実施例8
DL−カルニチンアミド塩酸塩20%(W/V)を含む
燐酸バッファー(pH7,0)501にカルニチンアミ
ド分解酵素生産菌を作用させるとL−カルニチンとD−
カルニチンアミドを得た。この反応生成液上清を酸処理
し生じた沈澱を除去したのち10mM酢酸アンモニウム
−酢1!ff1(pH1No)で平衡化させた強酸性陽
交換樹脂(ダイヤイオン 8K I P )1001へ
通液した。樹脂を水洗したのち2qbアンモニア水を通
液することにより該樹脂からL−カルニチンのみが溶離
できた。この画分を濃縮乾固させ、さらに真空乾燥基中
で乾燥することによ、9L−カルニチン分子内塩12k
Iiを得た。Example 8 When a carnitinamide degrading enzyme-producing bacterium was applied to 501 phosphate buffer (pH 7.0) containing 20% (W/V) DL-carnitinamide hydrochloride, L-carnitine and D-
Carnitinamide was obtained. This reaction product supernatant was treated with acid to remove the resulting precipitate, and then 10mM ammonium acetate-vinegar 1! The solution was passed through a strongly acidic positive exchange resin (Diaion 8K I P ) 1001 equilibrated with ff1 (pH 1 No.). After washing the resin with water, only L-carnitine could be eluted from the resin by passing 2 qb aqueous ammonia through the resin. This fraction was concentrated to dryness and further dried in a vacuum dryer to obtain 9L-carnitine inner salt 12k.
I got Ii.
(回収率90僑)
[”l”n =2 a 6 (a =10 e Ht
O)m、p、 =195〜205 (dec )カ
ルニチン純度=100%(apbc)5体比率 =
92%(HPL()Y11脂に吸看残存したD−カル
ニチンアミドは8%酢酸アンモニウム水溶液を通液する
ことによシ樹脂から溶離した。この画分を凝縮し析出塩
分を除去しのち稀塩酸を加えカルニチンアミド塩酸塩と
し再び@縮した。残留物にエタノールを加えるとカルニ
チンアミド塩酸塩の粗結晶が析出した。これを稀エタノ
ールから再結させることによfiD−カルニチンアミド
塩酸塩の結晶2.45kgを得た。(Recovery rate: 90 people) [“l”n = 2 a 6 (a = 10 e Ht
O) m, p, = 195-205 (dec) Carnitine purity = 100% (apbc) 5-body ratio =
D-carnitinamide remaining in the 92% (HPL()Y11 fat) was eluted from the resin by passing an 8% ammonium acetate aqueous solution through it. This fraction was condensed to remove the precipitated salt, and then diluted with dilute hydrochloric acid. was added to form carnitinamide hydrochloride and condensed again. When ethanol was added to the residue, crude crystals of carnitinamide hydrochloride were precipitated. By recrystallizing this from dilute ethanol, fiD-carnitinamide hydrochloride crystal 2 was obtained. .45 kg was obtained.
(回収率82%)
Ctx3背=+1 !>9 (0=10 、1!、O)
m、p、 =255〜240 (dec )カルニチ
ンアミド純度=94%(TlPLC)D体比率
=981 (HPLO)カルニチン混入率 くαo
1% (HPIIO)L−カル−チン混入率=100
9%
(DTNB法)
(ホ)発明の効果
上述のように、本発明によってカルニチンおよびカルニ
チンアミドを含有する水溶液から両化合物を効率的に分
離精製することが可能となった。(Recovery rate 82%) Ctx3 back = +1! >9 (0=10, 1!, O)
m, p, = 255-240 (dec) Carnitinamide purity = 94% (TlPLC) D-isomer ratio
=981 (HPLO) Carnitine contamination rate αo
1% (HPIIO) L-carcine contamination rate = 100
9% (DTNB Method) (e) Effects of the Invention As described above, the present invention has made it possible to efficiently separate and purify carnitine and carnitinamide from an aqueous solution containing both compounds.
べ何 中 山 清Beka Nakayama Kiyoshi
Claims (1)
H1〜5の塩類の水溶液で平衡化させた陽イオン交換樹
脂に接触させて、カルニチンおよびカルニチンアミドを
該樹脂に吸着させたのち、アンモニア水、アミンまたは
アンモニウム塩の水溶液で該樹脂を処理してカルニチン
のみを溶離し、カルニチンアミドを樹脂に吸着状態に残
存せしめることによりカルニチンとカルニチンアミドを
分離することを特徴とするカルニチンおよびカルニチン
アミドの分離精製法。 2、カルニチンアミドを吸着している陽イオン交換樹脂
を、アンモニウム塩、アミン、アミンの塩の何れかの水
溶液で処理することにより、カルニチンアミドを溶離回
収することを特徴とするカルニチンアミドの分離精製法
。[Claims] 1. A carnitine aqueous solution containing carnitinamide is
After contacting with a cation exchange resin equilibrated with an aqueous solution of salts of H1 to 5 to adsorb carnitine and carnitinamide onto the resin, the resin is treated with an aqueous solution of ammonia water, an amine or an ammonium salt. 1. A method for separating and purifying carnitine and carnitinamide, which comprises separating carnitine and carnitinamide by eluting only carnitine and leaving carnitinamide in an adsorbed state on a resin. 2. Separation and purification of carnitinamide, characterized in that carnitinamide is eluted and recovered by treating a cation exchange resin adsorbing carnitinamide with an aqueous solution of ammonium salt, amine, or amine salt. Law.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP63038690A JPH01213258A (en) | 1988-02-23 | 1988-02-23 | Purification of carnitine and carnitine amide |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP63038690A JPH01213258A (en) | 1988-02-23 | 1988-02-23 | Purification of carnitine and carnitine amide |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH01213258A true JPH01213258A (en) | 1989-08-28 |
Family
ID=12532295
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP63038690A Pending JPH01213258A (en) | 1988-02-23 | 1988-02-23 | Purification of carnitine and carnitine amide |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH01213258A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0722724A1 (en) * | 1995-01-20 | 1996-07-24 | Sigma-Tau Industrie Farmaceutiche Riunite S.p.A. | Use of L-carnitine and its derivatives for reducing the toxic effects of cyclosporin-A and others immunosuppressants |
-
1988
- 1988-02-23 JP JP63038690A patent/JPH01213258A/en active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0722724A1 (en) * | 1995-01-20 | 1996-07-24 | Sigma-Tau Industrie Farmaceutiche Riunite S.p.A. | Use of L-carnitine and its derivatives for reducing the toxic effects of cyclosporin-A and others immunosuppressants |
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