JPH0572919B2 - - Google Patents
Info
- Publication number
- JPH0572919B2 JPH0572919B2 JP19601985A JP19601985A JPH0572919B2 JP H0572919 B2 JPH0572919 B2 JP H0572919B2 JP 19601985 A JP19601985 A JP 19601985A JP 19601985 A JP19601985 A JP 19601985A JP H0572919 B2 JPH0572919 B2 JP H0572919B2
- Authority
- JP
- Japan
- Prior art keywords
- methylcobalamin
- lower aliphatic
- acetone
- water
- highly purified
- 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.)
- Expired - Fee Related
Links
- JEWJRMKHSMTXPP-BYFNXCQMSA-M methylcobalamin Chemical compound C[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O JEWJRMKHSMTXPP-BYFNXCQMSA-M 0.000 claims description 37
- 235000007672 methylcobalamin Nutrition 0.000 claims description 37
- 239000011585 methylcobalamin Substances 0.000 claims description 37
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 33
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 22
- RMRCNWBMXRMIRW-BYFNXCQMSA-M cyanocobalamin Chemical compound N#C[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O RMRCNWBMXRMIRW-BYFNXCQMSA-M 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 17
- 238000004519 manufacturing process Methods 0.000 claims description 13
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 235000000639 cyanocobalamin Nutrition 0.000 claims description 10
- 239000011666 cyanocobalamin Substances 0.000 claims description 10
- 229960002104 cyanocobalamin Drugs 0.000 claims description 10
- 125000001931 aliphatic group Chemical group 0.000 claims description 9
- 239000011541 reaction mixture Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000000741 silica gel Substances 0.000 claims description 5
- 229910002027 silica gel Inorganic materials 0.000 claims description 5
- 239000003463 adsorbent Substances 0.000 claims description 4
- 239000007858 starting material Substances 0.000 claims description 3
- 238000007796 conventional method Methods 0.000 claims description 2
- AAMWFMOHRYFSEU-UHFFFAOYSA-N methanol;propan-2-one;hydrate Chemical compound O.OC.CC(C)=O AAMWFMOHRYFSEU-UHFFFAOYSA-N 0.000 claims description 2
- RXKJFZQQPQGTFL-UHFFFAOYSA-N dihydroxyacetone Chemical compound OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 claims 6
- DNIAPMSPPWPWGF-GSVOUGTGSA-N (R)-(-)-Propylene glycol Chemical compound C[C@@H](O)CO DNIAPMSPPWPWGF-GSVOUGTGSA-N 0.000 claims 4
- 239000000243 solution Substances 0.000 description 12
- -1 aliphatic ketones Chemical class 0.000 description 10
- YOZNUFWCRFCGIH-BYFNXCQMSA-L hydroxocobalamin Chemical compound O[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O YOZNUFWCRFCGIH-BYFNXCQMSA-L 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 238000001179 sorption measurement Methods 0.000 description 5
- 239000013076 target substance Substances 0.000 description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 235000002639 sodium chloride Nutrition 0.000 description 4
- 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 3
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 235000004867 hydroxocobalamin Nutrition 0.000 description 3
- 239000011704 hydroxocobalamin Substances 0.000 description 3
- 229960001103 hydroxocobalamin Drugs 0.000 description 3
- 239000003456 ion exchange resin Substances 0.000 description 3
- 229920003303 ion-exchange polymer Polymers 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000002156 adsorbate Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002452 interceptive effect Effects 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 238000007069 methylation reaction Methods 0.000 description 2
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical compound O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 206010002065 Anaemia megaloblastic Diseases 0.000 description 1
- 241000694440 Colpidium aqueous Species 0.000 description 1
- 208000032131 Diabetic Neuropathies Diseases 0.000 description 1
- 208000000682 Megaloblastic Anemia Diseases 0.000 description 1
- 206010029240 Neuritis Diseases 0.000 description 1
- 206010036105 Polyneuropathy Diseases 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical group [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 231100001016 megaloblastic anemia Toxicity 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 208000027232 peripheral nervous system disease Diseases 0.000 description 1
- 208000033808 peripheral neuropathy Diseases 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 208000019629 polyneuritis Diseases 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000022558 protein metabolic process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000006276 transfer reaction Methods 0.000 description 1
Landscapes
- Nitrogen Condensed Heterocyclic Rings (AREA)
- Saccharide Compounds (AREA)
Description
【発明の詳細な説明】
産業上の利用分野
本発明は、医薬として実用に供しうる高純度の
メチルコバラミンの製造方法に関する。DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a method for producing highly purified methylcobalamin that can be practically used as a medicine.
従来の技術
メチルコバラミンは、血液および髄液存在型の
補酵素型B12として、神経細胞などの組織移行性
に優れている。生化学的にはメチル基転移反応に
関与し、核酸・蛋白・脂質代謝においてB12同族
体の中で最も優れた生体内活性を示す。臨床的に
は、巨赤芽球性貧血に有効であるのみならず、糖
尿病性神経障害、多発性神経炎などの末梢性神経
障害治療剤として有効である。Background Art Methylcobalamin, as coenzyme type B12 present in blood and cerebrospinal fluid, has excellent migration properties into tissues such as nerve cells. Biochemically, it is involved in methyl group transfer reactions, and exhibits the highest in vivo activity among B12 homologs in nucleic acid, protein, and lipid metabolism. Clinically, it is effective not only for megaloblastic anemia but also as a therapeutic agent for peripheral neuropathies such as diabetic neuropathy and polyneuritis.
メチルコバラミンの製法についての種々の方法
が知られているが、代表的な方法としては例えば
出発原料としてシアノコバラミンを使用する方法
がある（特公昭45−38059）。 Various methods for producing methylcobalamin are known, and a typical method is, for example, a method using cyanocobalamin as a starting material (Japanese Patent Publication No. 38059/1983).
発明が解決しようとする問題点
この合成法によつて得られたメチルコバラミン
含有溶液中には、微量のシアノコバラミン、ヒド
ロキソコバラミン、更に黄色物質、塩類などが含
有されている。したがつて、いかにしてこれらの
メチルコバラミン以外の挾雑成分を除き、高純度
のメチルコバラミンを得るかが問題である。Problems to be Solved by the Invention The methylcobalamin-containing solution obtained by this synthesis method contains trace amounts of cyanocobalamin, hydroxocobalamin, yellow substances, salts, and the like. Therefore, the problem is how to remove these interfering components other than methylcobalamin and obtain highly pure methylcobalamin.
これらの挾雑成分を除去する方法としては、こ
れらの物質は塩類を除いてその性質が近似してい
るため、吸着性の差を利用した方法、例えば、イ
オン交換樹脂クロマトグラフイーによる方法およ
び水−アセトン結晶化法などがすでに知られてい
る。 Methods for removing these interfering components include methods that take advantage of differences in adsorption properties, such as methods using ion exchange resin chromatography and water -Acetone crystallization methods are already known.
しかしながら、これらの方法は、次のような欠
点を有する。すなわち、イオン交換樹脂交脂クロ
マトグラフイーによる方法は、イオン交換樹脂が
高価なため、再利用するための別途設備を要する
という欠点を有し、また水−アセトン結晶化法で
は化学的性質が近似しているため、メチルコバラ
ミンの分離精製は困難であるという欠点を有して
いる。 However, these methods have the following drawbacks. That is, the method using ion-exchange resin crosslinking chromatography has the disadvantage that the ion-exchange resin is expensive and requires separate equipment for reuse, and the water-acetone crystallization method has the disadvantage that the chemical properties are not similar. Therefore, it has the disadvantage that it is difficult to separate and purify methylcobalamin.
問題点を解決するための手段
そこで、本発明者等は、これらの欠点を有しな
い高純度メチルコバラミンを得るため、長年にわ
たつて研究を重ねてきたが、次の手段により解決
できることを見い出し、本発明を完成した。すな
わち、
シアノコバラミンを常法によりメチル化して、
メチルコバラミンを主成分とする反応混合物を
得、この反応混合物を出発物質として、これを精
製し、高純度のメチルコバラミンを得る工程にお
いて、次の４つの工程を結合することを特徴とす
る高純度のメチルコバラミンの製造方法。Means for Solving the Problems Therefore, the present inventors have conducted research for many years in order to obtain high purity methylcobalamin that does not have these drawbacks, and have found that the problem can be solved by the following means. The invention has been completed. That is, cyanocobalamin is methylated by a conventional method,
A high-purity method characterized by combining the following four steps in the process of obtaining a reaction mixture containing methylcobalamin as a main component, and purifying this reaction mixture as a starting material to obtain high-purity methylcobalamin. A method for producing methylcobalamin.
反応混合液のPHを約2.5〜7.0にする工程、
シリカゲルに接触させ、該吸着物に低級脂肪
族ケトン−水を流出せしめる工程
該吸着物に、低級脂肪族アルコール類−水を
流出せしめ、吸着物を留出する工程
該留出液を濃縮し、低級脂肪族アルコール類
−水−低級脂肪族ケトン類に溶解し、該溶液に
低級脂肪族ケトン類を加えて結晶化させ、結晶
部より高純度のメチルコバラミンを得る工程
の工程
反応混合物のPHを約2.5〜7.0に調整する工程で
あるが、通常は希塩酸水溶液、好ましくは0.03N
以下の希塩酸を用いておこなう。PHは５前後が最
も好ましい。この範囲のPHに調整することによつ
て吸着量の増加と溶出分離に著しく効果がある。
PH7.0を超えた場合は未分離となり不利である。 A step of adjusting the pH of the reaction mixture to about 2.5 to 7.0. A step of bringing the reaction mixture into contact with silica gel and causing the lower aliphatic ketone-water to flow out onto the adsorbent. A step of causing the lower aliphatic alcohol-water to flow out into the adsorbent and adsorption. Step of distilling a substance: Concentrate the distillate, dissolve it in lower aliphatic alcohol-water-lower aliphatic ketones, add lower aliphatic ketones to the solution and crystallize it, Step of obtaining pure methylcobalamin This is a step of adjusting the pH of the reaction mixture to approximately 2.5 to 7.0, usually using a dilute aqueous hydrochloric acid solution, preferably 0.03N.
This is done using the following dilute hydrochloric acid. The pH is most preferably around 5. Adjusting the pH within this range has a significant effect on increasing adsorption amount and elution separation.
If the pH exceeds 7.0, it becomes unseparated and is disadvantageous.
の工程
このメチル化反応液を、シリカゲルに接触させ
る工程である。シリカゲルとしては例えばシリカ
ゲル−40（メルク社製）などを用いる。シリカゲ
ル−40は細孔径が40Åで他のシリカゲルに比し大
きく吸着に有利である。 This is the step of bringing this methylation reaction solution into contact with silica gel. As the silica gel, for example, silica gel-40 (manufactured by Merck & Co., Ltd.) is used. Silica gel-40 has a pore diameter of 40 Å, which is larger than other silica gels and is advantageous for adsorption.
溶出は、低級脂肪族ケトン−水、好ましくはア
セトンを用い、sv0.5〜1.0でおこなうことが好結
果を与える。アセトンは、約５％のアセトンルが
好ましい。10％以上のアセトン水溶液では未分離
で、全成分が溶出する可能性がある。 Elution using lower aliphatic ketone-water, preferably acetone, at sv 0.5 to 1.0 gives good results. Preferably, the acetone is about 5% acetone. In an aqueous solution of 10% or more acetone, all components may be eluted without being separated.
この操作により目的物質は吸着部にあり、コバ
ルト原子にメチル基が２個配位した副生物塩化ナ
トリウムなどが非吸着部分として除かれる。 By this operation, the target substance is present in the adsorption part, and by-products such as sodium chloride, in which two methyl groups are coordinated to a cobalt atom, are removed as non-adsorbed parts.
の工程
目的物質であるメチルコバラミンを含む吸着物
に、更に低級脂肪族アルコール類−水を流出せし
め、吸着物を留出する工程である。この際svは
0.5〜1.0の範囲でおこなうことが好ましい。 This step is a step in which lower aliphatic alcohol-water is further allowed to flow out into the adsorbate containing methylcobalamin, which is the target substance, and the adsorbate is distilled out. At this time, sv is
It is preferable to do this in the range of 0.5 to 1.0.
低級脂肪族アルコール類とは、メタノール、エ
タノール、プロパノール、ブタノールなどがあげ
られるが、メタノールが最も好ましい。メタノー
ルの濃度は15％前後が好ましい結果を与える。 Examples of lower aliphatic alcohols include methanol, ethanol, propanol, butanol, and methanol is most preferred. A methanol concentration of around 15% gives preferable results.
この工程により目的物質であるメチルコバラミ
ンは非吸着部分にあり、吸着されるのは黄色不純
物およびヒドロキソコバラミンである。非吸着部
分には、メチルコバラミンのほか、シアノコバラ
ミンが存在するものと考えられる。 Through this process, the target substance methylcobalamin is present in the non-adsorbed portion, and what is adsorbed are yellow impurities and hydroxocobalamin. It is thought that cyanocobalamin is present in the non-adsorbed portion in addition to methylcobalamin.
の工程
非吸着部分に含まれている目的物質であるメチ
ルコバラミンを取り出す工程であり、本工程によ
つて高純度メチルコバラミンが得られる。 This step is a step of removing the target substance methylcobalamin contained in the non-adsorbed portion, and high purity methylcobalamin is obtained through this step.
具体的には、の工程により得られた留出液
（非吸着部分）を濃縮した後、低級脂肪族アルコ
ール類−水−低級脂肪族ケトン類に溶解し、次い
で該溶液に低級脂肪族ケトン類を加えて結晶化さ
せ、結晶部より高純度のメチルコバラミンを得
る。 Specifically, after concentrating the distillate (non-adsorbed portion) obtained in the step, it is dissolved in lower aliphatic alcohol-water-lower aliphatic ketones, and then lower aliphatic ketones are added to the solution. is added to crystallize, and highly purified methylcobalamin is obtained from the crystallized portion.
低級脂肪族アルコールとは、メタノール、エタ
ノール、プロパノール、ブタノールなどをいい、
低級脂肪族ケトン類とは、アセトン、メチルエチ
ルケトンなどをいう。これらのうち、最も好まし
いものは、メタノールとアセトンである。メタノ
ール−水−アセトン等の最も好ましい割合は、約
2.5：1.10である。 Lower aliphatic alcohols include methanol, ethanol, propanol, butanol, etc.
Lower aliphatic ketones include acetone, methyl ethyl ketone, and the like. Among these, the most preferred are methanol and acetone. The most preferred ratio of methanol-water-acetone etc. is about
2.5:1.10.
本工程は、留出液に含まれている微量のシアノ
コバラミンと、目的物質であるメチルコバラミン
とを分離する方法であり、シアノコバラミンとメ
チルコバラミンの溶解差を利用している。 This process is a method for separating a trace amount of cyanocobalamin contained in the distillate from the target substance methylcobalamin, and utilizes the solubility difference between cyanocobalamin and methylcobalamin.
本発明のメリツトを要約すると次のとおりであ
る。 The advantages of the present invention are summarized as follows.
(1) 高純度・高収率のメチルコバラミンを得る工
業的な方法である。(1) It is an industrial method for obtaining methylcobalamin with high purity and high yield.
(2) 本法では、シリカゲルクロマトの再精製工程
を、低級アルコールおよび低級ケトンの水溶液
で同一カラム内での溶出除去をおこなうため、
大幅な時間短縮と、精製設備が不要となる。(2) In this method, the repurification step of silica gel chromatography is performed by elution and removal in the same column with an aqueous solution of lower alcohol and lower ketone.
Significant time savings and no need for purification equipment.
次に、本発明の代表的な実施例を掲げるが、本
発明がこれらのみに限定されることがないことは
いうまでもない。 Next, typical examples of the present invention will be listed, but it goes without saying that the present invention is not limited to these only.
実施例 １
特公昭45−38059に記載されているシアノコバ
ラミンからメチルコバラミンを製造する方法にし
たがい還元・メチル化反応（シアノコバラミン８
ｇ）をおこない、反応溶液から残余のヨードメチ
ルを濃縮留去し、95％メチルコバラミン含有メタ
ノール水溶液600c.c.を得た。この溶液を希塩酸水
溶液（0.03N以下）を徐々に滴下し、PH5.0とす
る。次いでシリカゲル（Kieselgel−40 メルク
社製）500c.c.が充填されたカラムにsv0.5の流速で
吸着させる。次いで５％アセトン水溶液600c.c.を
sv0.5の流速にて流し、ヒドロキソコバラミン、
食塩および金属錯体塩の除去をおこなう。その後
50％メタノール水溶液800c.c.をsv1.0の流速にてカ
ラムに流し、微量のシアノコバラミンを含有する
メチルコバラミン溶液500c.c.を溶出した。この溶
液をリン酸−セルロースおよびダウエツクス２
（Dowex２）よりなる混合床カラム（10c.c.）を
用いて、sv1.0の流速にて流し、通過液は60℃加
温下、減圧濃縮し、100c.c.水溶液とした。Example 1 Reduction and methylation reaction (cyanocobalamin 8
g) was carried out, and the remaining iodomethyl was distilled off from the reaction solution to obtain 600 c.c. of an aqueous methanol solution containing 95% methylcobalamin. A dilute aqueous hydrochloric acid solution (0.03N or less) is gradually added dropwise to this solution to adjust the pH to 5.0. Next, it is adsorbed onto a column packed with 500 c.c. of silica gel (Kieselgel-40 manufactured by Merck & Co.) at a flow rate of sv0.5. Next, add 600 c.c. of 5% acetone aqueous solution.
Flowed at a flow rate of sv0.5, hydroxocobalamin,
Removes common salt and metal complex salts. after that
800 c.c. of a 50% aqueous methanol solution was passed through the column at a flow rate of sv1.0, and 500 c.c. of a methylcobalamin solution containing a trace amount of cyanocobalamin was eluted. This solution was mixed with phosphoric acid-cellulose and Dowex 2.
A mixed bed column (10 c.c.) made of (Dowex 2) was used to flow at a flow rate of sv1.0, and the permeate was concentrated under reduced pressure while heating at 60° C. to form a 100 c.c. aqueous solution.
次にメタノール250c.c.を加え、40℃１時間溶解
後冷却下アセトン1000c.c.を徐々に滴下し、結晶
7.0ｇ（収率87.5％）を得た。 Next, add 250 c.c. of methanol, dissolve at 40℃ for 1 hour, and gradually add 1000 c.c. of acetone while cooling to crystallize.
7.0 g (yield 87.5%) was obtained.
結晶はHPLC法にて、メチルコバラミン含量
99.7％であり、シアノコバラミンは検出されなか
つた。 The content of crystals was determined by HPLC method.Methylcobalamin content
99.7%, and no cyanocobalamin was detected.
Claims (1)
て、メチルコバラミンを主成分とする反応混合物
を得、この反応混合物を出発物質として、これを
精製し、高純度のメチルコバラミンを得る工程に
おいて、次の４つの工程を結合することを特徴と
する高純度のメチルコバラミンの製造法。 反応混合液のPHを約2.5〜7.0に調整する工
程、 シリカゲルに接触させ、該吸着物に低級脂肪
族ケトン−水を流出させる工程、 次いで該吸着物に、低級脂肪族アルコール−
水を流出せしめ、吸着物を留出する工程、 該留出液を濃縮し、低級脂肪族アルコール−
水−低級脂肪族ケトンに溶解し、該溶液に低級
脂肪族ケトンを加えて結晶化させ、結晶部より
高純度のメチルコバラミンを得る工程。 ２ 工程において、流速svが0.5〜1.0である特
許請求の範囲第１項記載の高純度のメチルコバラ
ミンの製造方法。 ３ 工程において、流速svが0.5〜1.0である特
許請求の範囲第１項記載の高純度のメチルコバラ
ミンの製造方法。 ４ 工程およびにおいて、流速svが0.5〜1.0
である特許請求の範囲第１項記載の高純度のメチ
ルコバラミンの製造方法。 ５ 工程およびにおいて、低級脂肪族ケトン
がアセトンである特許請求の範囲第１項記載の高
純度のメチルコバラミンの製造方法。 ６ 工程およびにおいて、低級脂肪族アルコ
ールがメタノールである特許請求の範囲第１項記
載の高純度のメチルコバラミンの製造方法。 ７ 工程において、低級脂肪族アルコール−水
−低級脂肪族ケトンが2.5：１：10である特許請
求の範囲第１項記載の高純度のメチルコバラミン
の製造方法。 ８ 工程において、低級脂肪族アルコールがメ
タノールであり、低級脂肪族ケトンがアセトンで
ある特許請求の範囲第７項記載の高純度のメチル
コバラミンの製造方法。 ９ 工程において、PHが5.0〜6.0である特許請
求の範囲第１項記載の高純度のメチルコバラミン
の製造方法。 １０ 工程において、PHが5.0〜6.0であり、工
程およびにおいて、低級脂肪族ケトンがアセ
トンであり、工程およびにおいて、低級脂肪
族アルコールがメタノールであり、工程および
において、流速svが0.5〜1.0であり、工程に
おけるメタノール−水−アセトンが2.5：１：10
の割合である特許請求の範囲第１項記載の高純度
のメチルコバラミンの製造方法。[Claims] 1. A step in which cyanocobalamin is methylated by a conventional method to obtain a reaction mixture containing methylcobalamin as a main component, and this reaction mixture is used as a starting material and purified to obtain highly pure methylcobalamin. A method for producing high purity methylcobalamin, which is characterized by combining the following four steps. A step of adjusting the pH of the reaction mixture to about 2.5 to 7.0, a step of bringing it into contact with silica gel and allowing the lower aliphatic ketone-water to flow out onto the adsorbent, and then adding a lower aliphatic alcohol to the adsorbent.
A step of draining water and distilling off the adsorbed matter, concentrating the distillate and lower aliphatic alcohol-
A step of dissolving in water-lower aliphatic ketone, adding lower aliphatic ketone to the solution and crystallizing it, and obtaining highly purified methylcobalamin from the crystallized portion. 2. The method for producing highly purified methylcobalamin according to claim 1, wherein the flow rate sv in step 2 is 0.5 to 1.0. 3. The method for producing highly purified methylcobalamin according to claim 1, wherein the flow rate sv is 0.5 to 1.0 in step 3. 4 In the process, the flow rate sv is 0.5 to 1.0
A method for producing highly purified methylcobalamin according to claim 1. 5. The method for producing high-purity methylcobalamin according to claim 1, wherein the lower aliphatic ketone in step 5 is acetone. 6. The method for producing high-purity methylcobalamin according to claim 1, wherein the lower aliphatic alcohol in step 6 is methanol. 7. The method for producing high purity methylcobalamin according to claim 1, wherein in step 7, the ratio of lower aliphatic alcohol-water-lower aliphatic ketone is 2.5:1:10. 8. The method for producing high purity methylcobalamin according to claim 7, wherein in step 8, the lower aliphatic alcohol is methanol and the lower aliphatic ketone is acetone. 9. The method for producing highly purified methylcobalamin according to claim 1, wherein in step 9, the pH is 5.0 to 6.0. 10 In the step, the PH is 5.0 to 6.0, in the step and, the lower aliphatic ketone is acetone, in the step and, the lower aliphatic alcohol is methanol, and in the step and, the flow rate sv is 0.5 to 1.0. , methanol-water-acetone in the process was 2.5:1:10
A method for producing highly purified methylcobalamin according to claim 1, wherein the proportion is as follows.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19601985A JPS6256490A (en) | 1985-09-06 | 1985-09-06 | Production on high-purity methylcobalamin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19601985A JPS6256490A (en) | 1985-09-06 | 1985-09-06 | Production on high-purity methylcobalamin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6256490A JPS6256490A (en) | 1987-03-12 |
| JPH0572919B2 true JPH0572919B2 (en) | 1993-10-13 |
Family
ID=16350868
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP19601985A Granted JPS6256490A (en) | 1985-09-06 | 1985-09-06 | Production on high-purity methylcobalamin |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6256490A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012165934A1 (en) * | 2011-05-30 | 2012-12-06 | Interquim, S.A. De C.V. | Methylcobalamin synthesis process |
-
1985
- 1985-09-06 JP JP19601985A patent/JPS6256490A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6256490A (en) | 1987-03-12 |
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