JPS60181096A - Purification of bile acid - Google Patents
Purification of bile acidInfo
- Publication number
- JPS60181096A JPS60181096A JP3536084A JP3536084A JPS60181096A JP S60181096 A JPS60181096 A JP S60181096A JP 3536084 A JP3536084 A JP 3536084A JP 3536084 A JP3536084 A JP 3536084A JP S60181096 A JPS60181096 A JP S60181096A
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- Prior art keywords
- acid
- aqueous solution
- bile
- hydroxide
- bile acid
- 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.)
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Abstract
Description
【発明の詳細な説明】
本発明はウルンデオキシコール酸、ケノデオキシコール
酸1 fc ハ3α−ヒドロキシ−7一ヶトコラン酸の
精製方法に関する。さらに詳しくは、それら胆汁酸中に
不純物として含まれるリトコール酸に代表されるモノヒ
ドロキシ胆汁酸(ただし。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for purifying ulundeoxycholic acid, chenodeoxycholic acid 1 fc 3α-hydroxy-7 monotocholanic acid. More specifically, monohydroxy bile acids, typified by lithocholic acid, which are contained as impurities in these bile acids (however,
3α−ヒドロキシ−7−ケトコラン酸を除く。以下同様
。)およびコラン酸に代表されるヒドロキシ基を持たな
い胆汁酸(以下、ノンヒドロキシ胆汁酸という。)等を
選択的に除去する方法に関する。Excluding 3α-hydroxy-7-ketocholanic acid. Same below. ) and bile acids without hydroxyl groups (hereinafter referred to as non-hydroxy bile acids) such as colanic acid.
ウルソデオキシコール酸およびケノデオキシコール酸は
胆石溶解剤、利胆剤として有用であシ。Ursodeoxycholic acid and chenodeoxycholic acid are useful as gallstone dissolving agents and choleretic agents.
3α−ヒドロキシ−7−ケトコラン酸はこれらの製造中
間体として有用である。ウルンデオキシコール酸、ケノ
デオキシコール酸および3αイヒドロキシ−7−ケトコ
ラン酸は1例えばコール酸(3α、7α、12α−トリ
ヒドロキシ−5β−コラン酸)を出発原料として製造し
、その際それぞれの製造工程に由来する副生成物1例え
ばリトコールp(3α−ヒドロキシ−5β−フラン酸)
。3α-hydroxy-7-ketocholanic acid is useful as an intermediate in their production. Urundeoxycholic acid, chenodeoxycholic acid, and 3α-trihydroxy-7-ketocholanic acid are produced using, for example, cholic acid (3α, 7α, 12α-trihydroxy-5β-cholanic acid) as a starting material. By-products derived from 1 such as lithol p (3α-hydroxy-5β-furanic acid)
.
コラン酸、7α(または7β)−ヒドロキシコラン酸、
3α、7α−ジヒドロキ7−12−ケトコラン酸および
未反応コール酸などが残存し、それらの分離精製が非常
に困難とされてきた。特にリトコール酸は好ましくない
毒性を有している・ため効率よく除去することが望まれ
ている。colanic acid, 7α (or 7β)-hydroxycholanic acid,
3α,7α-dihydroxy-7-12-ketocholanic acid and unreacted cholic acid remain, making it extremely difficult to separate and purify them. In particular, lithocholic acid has undesirable toxicity, so it is desired to remove it efficiently.
従来公知のウルソデオキシコール酸またはケノデオキシ
コール酸の精製方法を大別すると■エステル化またはア
シル化して再結晶あるいはカラムクロマトグラフィーを
おこなう方法(特開昭52−153954.同52−1
53955.同52−’156851)、■有機溶剤か
ら直接再結晶する方法(化学実験学才10巻、495頁
、1943年、特開昭52−139053.同53−1
″37945、同55−79398.同56−3249
8)、■アルカリ金属を用いる方法(特開昭49−95
(J 5s、同5O−126654)などである。し
かし、とiらは操作の簡便さ、収率、N製純度などの点
で必ずしも満足されるものではなかった。また1本発明
者らは先に粗製胆汁酸の精製法として、水に溶けにくい
有機溶剤とアルカリ水溶液を用いる二相抽出法による操
作方法について出願したが(特開昭58−113202
)、さらに研究を行なった結果、−モノヒドロキシ胆汁
酸およびノンヒドロキシ胆汁酸(これらは環内に。Conventionally known purification methods for ursodeoxycholic acid or chenodeoxycholic acid can be roughly divided into: 1. Methods of esterification or acylation followed by recrystallization or column chromatography (JP-A No. 52-153954. No. 52-1)
53955. 52-'156851), ■Direct recrystallization method from organic solvent (Kagaku Jikken Gakusai Vol. 10, p. 495, 1943, JP-A-52-139053. 53-1)
″37945, 55-79398. 56-3249
8), ■Method using alkali metals (Japanese Unexamined Patent Publication No. 1986-1995)
(J 5s, 5O-126654), etc. However, they were not necessarily satisfactory in terms of ease of operation, yield, purity of N product, etc. In addition, the present inventors had previously filed an application for a two-phase extraction method using an organic solvent that is hardly soluble in water and an alkaline aqueous solution as a purification method for crude bile acids (Japanese Patent Laid-Open No. 113-202-1982).
), further studies revealed that -monohydroxy bile acids and non-hydroxy bile acids (these are within the ring).
二重結合を有するものも含む。)を選択的、特異的かつ
効率よく分離除去する方法を見出だし、ここに発明を完
成した。Also includes those with double bonds. ) has been discovered, and the invention has now been completed.
モノヒドロキシ胆汁酸としてはりトコール酸。Tocholic acid as a monohydroxy bile acid.
7α−ヒドロキシコラン酸、7β−ヒドロキシコラン酸
、3β−ヒドロキシコラン酸等が挙げられ。Examples include 7α-hydroxycholanic acid, 7β-hydroxycholanic acid, 3β-hydroxycholanic acid, and the like.
ノンヒドロキシ胆汁酸としてはフラン酸等であり。Examples of non-hydroxy bile acids include furanic acid.
その池水酸基が脱離して生成する環内に二重結合を有す
る胆汁酸も含む(以下、これらを不純物胆汁酸と総称す
る。)。これらの不純物胆汁酸は単独または2種以上混
在していてもよい。不純物胆汁酸のうちリトコール酸は
毒性を有しており、またケノデオキシコール酸またはウ
ルソデオキシコール酸の通常の製造工程においてリトコ
ール酸が1チ前後副生ずることが避は難いので、これを
01係以下に効率よく除く精製方法が望まれている。It also includes bile acids that have a double bond in the ring produced by the elimination of the hydroxyl group (hereinafter, these are collectively referred to as impurity bile acids). These impurity bile acids may be used alone or in combination of two or more. Among the impurity bile acids, lithocholic acid is toxic, and in the normal manufacturing process of chenodeoxycholic acid or ursodeoxycholic acid, it is unavoidable that about 1 liter of lithocholic acid is produced as a by-product. A purification method that efficiently removes it is desired.
本発明はりトコール酸をはじめ上記の不純物胆汁酸を選
択的、特異的に除去する方法を提供する。The present invention provides a method for selectively and specifically removing the above-mentioned impurity bile acids including tocholic acid.
次に9本発明方法を説明する。Next, nine methods of the present invention will be explained.
リトコール酸、コラン酸等の不純物胆汁酸を含有する粗
製胆汁酸を10〜1.5倍モル量、好ましくは等モル量
のアルカリを含む水溶液に溶解し。A crude bile acid containing impurity bile acids such as lithocholic acid and colanic acid is dissolved in an aqueous solution containing an alkali of 10 to 1.5 times the molar amount, preferably an equimolar amount.
この水溶液中に、不純物胆汁酸に対し10〜150倍モ
ル量の下式で示される第4級アンモニウム塩0
3
〔式中R工、 R,、R,および鳥は同一または異なつ
て炭素数1〜30個のアルキル基またはアラルキル基を
示しくただし、R□、 R,、’R3,R4の水酸基ま
たはハロゲン原子を示す。〕
及び水と層分離し得る有機溶剤を加え、得られた混合液
を5〜10分間撹拌する。液が2層に分離するまで静置
したのち水層を分取する。この操作によって、はじめア
ルカリ塩を形成していた不純物胆汁酸は疎水性の第4級
アンモニウムと塩の交換を受け、第4級アンモニウム塩
と疎水性のイオン対化合物を優先的に形成するので有機
溶媒層に選択的に移行し水層艇ら除去される。粗製胆汁
酸の純度に応じてこの操作を1〜3回行う。有機溶剤と
アルカリ水溶液の量は0.8:1.0〜1.0 : 9
.0(v/v)が好ましい。分取した水溶液中に微量の
有機溶剤が残存するときは必要に応じて蒸溜によシ除去
する。次いで、水溶液中に希塩酸、希硫酸などの酸を加
えて酸性化すると目的胆汁酸が沈殿する。In this aqueous solution, a quaternary ammonium salt represented by the following formula is present in an amount of 10 to 150 times the molar amount of the impurity bile acid. Indicates ~30 alkyl groups or aralkyl groups, provided that R□, R,, 'R3, and R4 represent hydroxyl groups or halogen atoms. ] and an organic solvent that can be separated into layers from water, and the resulting mixture is stirred for 5 to 10 minutes. After allowing the liquid to stand until it separates into two layers, separate the aqueous layer. Through this operation, the impurity bile acid, which initially formed an alkaline salt, undergoes a salt exchange with the hydrophobic quaternary ammonium, preferentially forming a hydrophobic ion pair compound with the quaternary ammonium salt. It selectively transfers to the solvent layer and is removed from the water layer. This operation is carried out 1 to 3 times depending on the purity of the crude bile acid. The amount of organic solvent and alkaline aqueous solution is 0.8:1.0 to 1.0:9
.. 0 (v/v) is preferred. If a trace amount of organic solvent remains in the separated aqueous solution, it is removed by distillation if necessary. Next, when the aqueous solution is acidified by adding an acid such as dilute hydrochloric acid or dilute sulfuric acid, the target bile acid is precipitated.
本発明において使用されるアルカリとしては水酸化ナト
リウム、水酸化カリウム、水酸化アンモニウム等の水酸
化アルカリ、炭酸ナトリウム、炭酸カリウム、炭酸アン
モニウム等の炭酸アルカリ。The alkali used in the present invention includes alkali hydroxides such as sodium hydroxide, potassium hydroxide, and ammonium hydroxide, and alkali carbonates such as sodium carbonate, potassium carbonate, and ammonium carbonate.
モノエタノールアミン、ピペラジン、ピペリジン。Monoethanolamine, piperazine, piperidine.
ピリジン等の有機塩基が挙げられる。又、前記の式で示
される第4Rアンモニウム塩は、具体的に。Examples include organic bases such as pyridine. Further, the 4R ammonium salt represented by the above formula is specifically:
水酸化テトラプロピルアンモニウム、水酸化テトラブチ
ルアンモニウム、水酸化テトラペンチルアンモニウム、
水酸化テトラヘキシルアンモニウム。Tetrapropylammonium hydroxide, Tetrabutylammonium hydroxide, Tetrapentylammonium hydroxide,
Tetrahexylammonium hydroxide.
水酸化テトラヘプチルアンモニウム、水酸イヒトリオク
チルメチルアンモニウム、水酸化セチ”ルトリメチルア
ンモニウム、水酸化ベンジルトリメチルアンモニウム、
水酸化ベンジルトリエチルアンモニウム、水酸化7エネ
チルトリメチルアンモニウム、水酸化スチリルトリメチ
ルアンモニウム、水酸化l−ラウリルピリジニウム、水
酸化lラウリル−4−ピコリニウム、水酸化トリエイコ
シルメチルアンモニウム、水酸化テトラアコンチルアン
モニウム、塩化テトラヘプチルアンモニウム、塩化1−
2ウリル−4−ピコリウム、塩化ベンジルトリメチルア
ンモニウム、塩化ベンジルトリブチルアンモニウムなど
である。水と層分離し得る有機溶剤としてはn−ブタノ
ール、 5ec−ブタノニル。Tetraheptylammonium hydroxide, hytrioctylmethylammonium hydroxide, cetyltrimethylammonium hydroxide, benzyltrimethylammonium hydroxide,
Benzyltriethylammonium hydroxide, 7enethyltrimethylammonium hydroxide, styryltrimethylammonium hydroxide, l-laurylpyridinium hydroxide, llauryl-4-picolinium hydroxide, trieicosylmethylammonium hydroxide, tetraacontylammonium hydroxide , tetraheptyl ammonium chloride, 1-chloride
These include 2uryl-4-picolium, benzyltrimethylammonium chloride, benzyltributylammonium chloride, and the like. Examples of organic solvents that can be separated into layers from water include n-butanol and 5ec-butanonyl.
n−アミルアルコール、n−ヘキサン等のアルコール類
、酢酸エチル、酢酸ブチル等のエステル類。Alcohols such as n-amyl alcohol and n-hexane, and esters such as ethyl acetate and butyl acetate.
メチルイソブチルケトン等のケトン類、エチルエーテル
、イングロビルエーテル、n−ブチルエーテル、アニソ
ール、エチレンクリコールシフチルエーテル等のエーテ
ル類、クロロホルム、ジクロルエタン、四塩化炭素等の
ハロゲン化炭化水素類。Ketones such as methyl isobutyl ketone, ethers such as ethyl ether, inglovir ether, n-butyl ether, anisole, and ethylene glycol cyphthyl ether, and halogenated hydrocarbons such as chloroform, dichloroethane, and carbon tetrachloride.
ベンゼン、シクロヘキサン、n−ヘキサン等の炭化水素
類の単独または混合溶媒であり、混合溶媒として使用す
るのが好ましい。It is a single or mixed solvent of hydrocarbons such as benzene, cyclohexane, n-hexane, etc., and is preferably used as a mixed solvent.
以下に本発明の精製方法を実施例をもって説明する。た
だし、各実施例中の不純物胆汁酸の含量はガスクロマト
グラフィー(日立163型ガスクロマトグラフイー。内
部標準にフラン酸またはコール酸を使用。)及び薄層ク
ロマトグラフィー〔シリカゲル60(メルク社M)、展
開溶媒は酢酸エチル:シクロヘキサン;氷酢酸(50:
13:3)〕によって測定した。The purification method of the present invention will be explained below with reference to Examples. However, the content of impurity bile acid in each example was determined by gas chromatography (Hitachi Model 163 gas chromatography. Furanic acid or cholic acid was used as an internal standard) and thin layer chromatography [Silica gel 60 (Merck M), The developing solvent was ethyl acetate: cyclohexane; glacial acetic acid (50:
13:3)].
実施例 1
リトコール酸0.5%、7β−ヒドロキシコラン酸03
%、ケノデオキシコール酸10%を含有するウルソデオ
キシコール酸100tを水酸化ナトリウム10.2’r
(等モル量)を含有する水溶液31に溶解し、この水溶
液に水酸化トリオクチルメチルアンモニウム347グ(
純度85係、不純物胆汁酸に対して359倍モル量)及
びシクロヘキサン−5ec−ブタノール(8:2)の混
合解媒11を加え5分間撹拌した。液が2層に分離する
まで静置してから下層の水層を分取した。再び水層に水
酸化トリオクチルメチルアンモニウーム2311(不純
物胆汁酸に対して2.39倍モル量)及びシクロヘキサ
ン−5ec−ブタノール(8:2)の混合溶媒1eを加
えて前記と同様の操作を行い水層を分取した。次いで、
液量が2.71になるまで濃縮して水溶液中に微量に存
在する有機溶媒を除去しこの水溶液中にO9l規定硫酸
を滴下すると無定形固体が沈殿した。これをろ取し、水
洗し、乾燥してウルソデオキシコール酸94.5 It
’ (回収率94.5%)を得た。リトコール酸は0.
04%、7β−ヒドロキシコラン酸は001%以下に減
少し、ケノデオキシコール酸は10チであった。Example 1 Lithocholic acid 0.5%, 7β-hydroxycholanic acid 03
%, 100 t of ursodeoxycholic acid containing 10% of chenodeoxycholic acid was added to 10.2'r of sodium hydroxide.
(equimolar amount), and in this aqueous solution, 347 g of trioctylmethylammonium hydroxide (
A mixed decomposition solvent 11 of cyclohexane-5ec-butanol (8:2) and cyclohexane-5ec-butanol (8:2) was added and stirred for 5 minutes. The solution was allowed to stand until it separated into two layers, and then the lower aqueous layer was separated. A mixed solvent 1e of trioctylmethylammonium hydroxide 2311 (2.39 times the molar amount relative to the impurity bile acid) and cyclohexane-5ec-butanol (8:2) was added to the aqueous layer again, and the same operation as above was performed. The aqueous layer was separated. Then,
The solution was concentrated to a liquid volume of 2.71 to remove a trace amount of organic solvent present in the aqueous solution, and 09l normal sulfuric acid was dropped into the aqueous solution to precipitate an amorphous solid. This was collected by filtration, washed with water, and dried to give 94.5 It of ursodeoxycholic acid.
' (recovery rate of 94.5%) was obtained. Lithocholic acid is 0.
0.04%, 7β-hydroxycholanic acid decreased to less than 0.001%, and chenodeoxycholic acid decreased to 10%.
実施例 2
リトコール酸0.4%、7α−ヒドロキシコラン酸02
%、3α、7α−ジヒドロキシ−12−ケトコラン酸2
5%、コール酸0.5%を含有するケノデオキシコール
酸3007を水酸化カリウム45゜3f(等モル量)を
含有する水溶液91に溶゛解し。Example 2 Lithocholic acid 0.4%, 7α-hydroxycholanic acid 02
%, 3α, 7α-dihydroxy-12-ketocholanic acid 2
Chenodeoxycholic acid 3007 containing 5% and 0.5% cholic acid was dissolved in an aqueous solution 91 containing 45°3f (equimolar amount) of potassium hydroxide.
この水溶−に水酸化テトラペンチルアンモニウム4.8
3F、(不純物胆汁酸に対して3.20倍モル量)及び
クロロホルム−四塩化炭素(1: 1’ )の混合溶媒
11を加え5分間撹拌した。液が2層に分離するまで静
置してから上層の水層を分取した。In this aqueous solution, 4.8% tetrapentylammonium hydroxide was added.
3F (3.20 times the molar amount relative to the impurity bile acid) and a mixed solvent 11 of chloroform-carbon tetrachloride (1:1') were added and stirred for 5 minutes. The liquid was allowed to stand until it separated into two layers, and then the upper aqueous layer was separated.
さらに水酸化テトラペンチルアンモニウム2.41(不
純物胆汁酸に対して159倍モル量)を使用して前記と
同様の操作を2回行った。分取した水溶液を液量が84
になるまで減圧濃縮して水溶液中に微量存在する有機溶
媒を除去し、この水溶液中に0.1規定硫酸を滴下する
と無定形固体が沈殿した。これをろ取し、水洗し、乾燥
してケノデオキX7 コ−炭酸286.52(回収率9
5.5 % )を得た。リトコール酸は0.04%、7
α−ヒドロキシコラン酸は0.01%以下に減少し、3
α、7α−ジヒドロキシ−12−ケトコラン酸は2.5
%、 コール酸は0.5%であった。Furthermore, the same operation as above was carried out twice using 2.41 times the molar amount of tetrapentylammonium hydroxide (159 times the molar amount relative to the impurity bile acid). The volume of the separated aqueous solution is 84
The aqueous solution was concentrated under reduced pressure to remove a trace amount of organic solvent present in the aqueous solution, and 0.1N sulfuric acid was dropped into the aqueous solution to precipitate an amorphous solid. This was collected by filtration, washed with water, and dried to yield 286.52 (recovery rate of 9).
5.5%). Lithocholic acid is 0.04%, 7
α-hydroxycholanic acid decreased to less than 0.01%, 3
α,7α-dihydroxy-12-ketocholanic acid is 2.5
%, cholic acid was 0.5%.
実施例 3
リトコール酸4.0%、フラン酸2.0%を含有する3
α−ヒドロキシ−7−ケトコラン酸2fを炭酸ナトリウ
ム0.286 P (等モル量)を含有する水溶液50
WLeに溶解し、この水溶液に水酸化テトラヘプチール
ーアンモニウム11’4.2 rn:i (不純物11
jl酸に対して0824倍モル量)及び+1−ブチルエ
ーテル−〇−アミルアルコール(9:1)の混合′ 溶
媒25rrtlを加え5分間撹拌した。液が2層に分離
するまで静置してから下層の水層を分取した。Example 3 3 containing 4.0% lithocholic acid and 2.0% furanic acid
α-hydroxy-7-ketocholanic acid 2f in an aqueous solution containing 0.286 P (equimolar amount) of sodium carbonate 50
Tetraheptylammonium hydroxide 11'4.2 rn:i (impurity 11
25 rrtl of a mixed solvent of 0824 times molar amount relative to Jl acid) and +1-butyl ether-〇-amyl alcohol (9:1) were added and stirred for 5 minutes. The solution was allowed to stand until it separated into two layers, and then the lower aqueous layer was separated.
更に水層について前記と全く同様の操作を2回行い水層
を分取した。この水溶液中に0.1規定硫酸を滴下する
と無定形固体が沈殿し これをろ取し。Furthermore, the same operation as above was performed twice on the aqueous layer, and the aqueous layer was separated. When 0.1N sulfuric acid was added dropwise to this aqueous solution, an amorphous solid precipitated and was collected by filtration.
水洗し、乾燥して3α−ヒドロキシ−7−ケドコラン酸
1.70 j(回収率850%)を得た。−リトコール
酸は0.1%、フラン酸は0.05%以下に減少した。It was washed with water and dried to obtain 1.70 j of 3α-hydroxy-7-kedocholanic acid (recovery rate: 850%). -Lithocholic acid decreased to 0.1% and furanic acid decreased to 0.05% or less.
実施例 4
リトコール酸05%、7β−ヒドロキシコラン酸0.3
%、ケノデオキシコール酸10.0%を含有するウルソ
デオキシコール酸50!7’を水酸化ナトリウム7.6
6f(1,5倍モル量)を含有する水溶液1.3’ A
に溶解し、この水溶液に水酸化トリオクチルメチルアン
モニウム1.74 t (純度85%。Example 4 Lithocholic acid 05%, 7β-hydroxycholanic acid 0.3
50!7' of ursodeoxycholic acid containing 10.0% of chenodeoxycholic acid and 7.6% of sodium hydroxide.
Aqueous solution 1.3'A containing 6f (1.5 times the molar amount)
1.74 t of trioctylmethylammonium hydroxide (purity 85%) was dissolved in this aqueous solution.
不純物胆汁酸に対して3.60倍モル量)及びn−ヘキ
サン−〇−ブタノール(1:1)の混合溶媒05ぎを加
え5分間撹拌した。液が2層に分離するまで静置してか
ら下層の水層を分取した。再び水層へ水酸化トリオクチ
ルメチルアンモニウム1゜及び前記と同様の混合溶媒0
.57を加えて前記と同様の操作を行い水層を分取した
。分取した水溶液の液量が1.11になるまで減圧濃縮
して微量存在する有機溶媒を除去し、この水溶液中に0
1規定塩酸を滴下すると無定形固体が沈殿した。これを
ろ取し、水洗し、乾燥してウルソデオキシコール酸47
.251(回収率94.5 % )を得た。リトコール
酸は0.04%、7β−ヒドロキシコラン酸は0.01
%以下に減少しクツデオキシコール酸は9.9%であっ
た。A mixed solvent of 3.60 times the molar amount of impurity bile acid) and n-hexane-〇-butanol (1:1) was added and stirred for 5 minutes. The solution was allowed to stand until it separated into two layers, and then the lower aqueous layer was separated. Add 1° of trioctylmethylammonium hydroxide and 0% of the same mixed solvent as above to the aqueous layer again.
.. 57 was added and the same operation as above was performed to separate the aqueous layer. The fractionated aqueous solution is concentrated under reduced pressure until the liquid volume becomes 1.11 to remove a trace amount of organic solvent.
When 1N hydrochloric acid was added dropwise, an amorphous solid precipitated. This was collected by filtration, washed with water, and dried to produce ursodeoxycholic acid 47.
.. 251 (recovery rate 94.5%) was obtained. Lithocholic acid is 0.04%, 7β-hydroxycholanic acid is 0.01%
%, and cutudeoxycholic acid was 9.9%.
実施例 5
リトコール酸03%、7α−ヒドロキシコラン酸03%
を含有するケノデオキシコール酸2007をモノエタノ
ールアミン32842(等モル量)を含有する水溶液6
1に溶解し、この水溶液に塩化テトラヘプチルアンモニ
ウム4.369(不純物胆汁酸に対して307倍モル量
)及びクロロホルム−四塩化炭素(1:1)の混合溶媒
11を加え5分間撹拌した。液が2層に分離するまで静
置してから上層の水層を分取した。さらに水層に塩化テ
トラヘプチルアンモニウム2.187(不純物胆汁酸に
対して153倍モル量)及び前記と同様の混合溶媒11
を加えて前記と同様の操作を2回行った。分取した水溶
液を液量が51になるまで濃縮した後0.1規定硫酸を
滴下すると無定形固体が沈殿した。これをろ取し7゛水
洗し、乾燥してケノデオキシコール酸1’ 90. ’
O’ y (回収率95.0%)を得た。リトコール酸
は003%、7α−ヒドロキシコラン酸は0.01%以
下に減少した。Example 5 Lithocholic acid 03%, 7α-hydroxycholanic acid 03%
An aqueous solution 6 containing chenodeoxycholic acid 2007 containing monoethanolamine 32842 (equimolar amount)
A mixed solvent 11 of 4.369 tetraheptylammonium chloride (307 times the molar amount relative to the impurity bile acid) and chloroform-carbon tetrachloride (1:1) was added to this aqueous solution, and the mixture was stirred for 5 minutes. The liquid was allowed to stand until it separated into two layers, and then the upper aqueous layer was separated. Furthermore, in the aqueous layer, 2.187 tetraheptylammonium chloride (153 times the molar amount relative to the impurity bile acid) and the same mixed solvent 11 as above were added.
was added and the same operation as above was performed twice. After concentrating the separated aqueous solution to a liquid volume of 51, 0.1N sulfuric acid was added dropwise to precipitate an amorphous solid. This was collected by filtration, washed with water for 70 minutes, and dried to give chenodeoxycholic acid 1'90. '
O' y (recovery rate 95.0%) was obtained. Lithocholic acid decreased to 0.003%, and 7α-hydroxycholanic acid decreased to 0.01% or less.
実施例 6
リトコール酸05%、7β−ヒドロキシコラン酸0.3
%、ケノデオキシコール酸12%を含有するウルソデオ
キシコール酸107を水酸化ナトリウム1.029(等
モル量)を含有する水溶液300ゴに溶解し、この水溶
液に塩化1−ラウリル−4−ピコリニウムの30%水溶
液76 oq(不純物胆汁酸に対して3.59倍モル量
)及びクロロホルム10’OmJを加え5分間撹拌した
。液が2層に分離するまで静−置してから下層の水層を
分取した。Example 6 Lithocholic acid 05%, 7β-hydroxycholanic acid 0.3
%, ursodeoxycholic acid 107 containing chenodeoxycholic acid 12% is dissolved in 300 g of an aqueous solution containing 1.029 (equimolar amount) of sodium hydroxide, and 30% of 1-lauryl-4-picolinium chloride is dissolved in this aqueous solution. 76 oq of an aqueous solution (3.59 times the molar amount relative to the impurity bile acid) and 10'OmJ of chloroform were added and stirred for 5 minutes. The solution was allowed to stand until it separated into two layers, and then the lower aqueous layer was separated.
再び水層へ塩化1−ラウリル−4−ピコリニウムの30
%水溶液so7my(不純物胆汁酸に対して2.40倍
モル量)及びクロロホルム100meを加え、前記と同
様の操作を行い水層を分取した。次いで、液量が270
rrtlになるまで減圧濃縮したのち、水溶液中に0
.1規定塩酸を滴下すると無定形固体が沈殿した。これ
をろ取し、水洗し、乾燥してウルソデオキシコール酸9
47グ(回収率947%)を得た。リトコール酸は00
5%、7β−ヒドロキシコラン酸は001チ以下に減少
し、ケノデオキシコール酸は11%テアツタ。Add 30% of 1-lauryl-4-picolinium chloride to the aqueous layer again.
% aqueous solution so7my (2.40 times the molar amount relative to the impurity bile acid) and chloroform 100me were added, and the same operation as above was performed to separate the aqueous layer. Then, the liquid volume is 270
After concentrating under reduced pressure until it becomes rrtl, 0 is added to the aqueous solution.
.. When 1N hydrochloric acid was added dropwise, an amorphous solid precipitated. This was collected by filtration, washed with water, and dried to produce ursodeoxycholic acid 9.
47 g (recovery rate 947%) was obtained. Lithocholic acid is 00
5%, 7β-hydroxycholanic acid decreased to less than 001%, and chenodeoxycholic acid decreased to 11%.
実施例 7
リトコール酸0.3%、フラン酸0.3%を含有するケ
ノデオキシコール酸202を水酸化カリウム286り(
等モル量)を含有する水溶液600 rugに溶解し、
この水溶液に塩化ベンジルトリブチルアンモニウムの5
0%水溶液636■(不純物胆汁酸に対して3,13倍
モル量)及びクロロホルム−I+−ブタ/−ル(8:
2 )+17)混合溶媒100m/を加え5分間撹拌し
た。液が2層に分離するまで静置してから、上層の水層
を分取した。水層に対して前記と同様の操作を2回行い
水層を分取した。Example 7 Chenodeoxycholic acid 202 containing 0.3% lithocholic acid and 0.3% furanic acid was mixed with potassium hydroxide 286 (
equimolar amount) in an aqueous solution containing 600 rug of
Add 5% of benzyltributylammonium chloride to this aqueous solution.
0% aqueous solution 636 μm (3.13 times the molar amount relative to the impurity bile acid) and chloroform-I+-but/-ol (8:
2)+17) 100ml of mixed solvent was added and stirred for 5 minutes. After the liquid was allowed to stand until it separated into two layers, the upper aqueous layer was separated. The same operation as above was performed twice on the aqueous layer, and the aqueous layer was separated.
次いで、液量が550 mlになるまで濃縮して、水溶
液中に微量存在する有機溶媒を除去したのち。Next, the solution was concentrated to a volume of 550 ml to remove trace amounts of organic solvent present in the aqueous solution.
01規定硫酸を滴下すると無定形固体が沈殿した。When 0.1N sulfuric acid was added dropwise, an amorphous solid precipitated.
これをろ取し、水洗し、乾燥してケノデオキシコール酸
18.7r(回収率935%)を得た。リトコール酸は
0.06%、フラン酸は0.03%に減少した。This was collected by filtration, washed with water, and dried to obtain chenodeoxycholic acid 18.7r (recovery rate 935%). Lithocholic acid decreased to 0.06% and furanic acid decreased to 0.03%.
出 願 人 東京田辺製薬株式会社 代理人 久高将信(外−名) 手 続 補 正 ii(方式) 昭和59年6月19日 特許庁長官 若 杉 和 夫 殿 】、事件の表示 特 願 昭59−35,360 号 2発明の名称 胆汁酸の精製方法 3、補正をする者 事件との関係 特許出願人 東京田辺製薬株式会社 4代理人Applicant: Tokyo Tanabe Pharmaceutical Co., Ltd. Agent: Masanobu Hisataka (foreign name) Procedure correction ii (method) June 19, 1981 Mr. Kazuo Wakasugi, Commissioner of the Patent Office ], Incident display Special request No. 59-35,360 2. Name of the invention Method for purifying bile acids 3. Person who makes corrections Relationship to the incident: Patent applicant Tokyo Tanabe Pharmaceutical Co., Ltd. 4 agents
Claims (4)
カリ水溶液および下式で示される第4級アンモ3 〔式中几□、■2.■。およびR2は同一または異なっ
て炭素数1〜30個のアルキル基またはアラルキル基を
示しくただし、R1,〜、 R8,凡、の水酸基または
ハロゲン原子を示す。〕 を加えて撹拌したのち、水溶液層を分取し9分取した水
溶液に酸を加えて胆汁酸を沈殿させることを特徴とする
胆汁酸の精製方法。(1) An organic solvent capable of phase separation from water in the crude bile acid, an alkaline aqueous solution, and a quaternary ammonium 3 represented by the following formula [wherein the formula □, ■2. ■. and R2 are the same or different and represent an alkyl group or an aralkyl group having 1 to 30 carbon atoms, provided that R1, to R8, generally represent a hydroxyl group or a halogen atom. ] A method for purifying bile acids, which comprises adding and stirring, separating an aqueous solution layer, and adding an acid to nine aliquots of the aqueous solution to precipitate the bile acids.
ドコラン酸である特許請求の範囲オ(1)項記載の精製
方法。(2) The bile acid to be purified is ursodeoxycholic acid. The purification method according to claim E(1), which is chenodeoxycholic acid or 3α-hydroxy-7-kedocholanic acid.
ル酸、コラン酸、7α−ヒドロキシコラン酸または7β
−ヒドロキシコラン酸である特許請求の範囲オ(1)項
記載の精製方法。(3) Impurity bile acid contained in crude bile acid is lithocholic acid, colanic acid, 7α-hydroxycholanic acid or 7β
-Hydroxycholanic acid.
ルアンモニウム、水酸化テトラペンチルアンモニウム、
水酸化テトラヘプチルアンモニウムまたは塩化テトラヘ
プチルアンモニウムである特許請求の範囲オ(1)項の
記載の精製方法。(4) The quaternary ammonium salt is trioctylmethylammonium hydroxide, tetrapentylammonium hydroxide,
The purification method according to claim E(1), which is tetraheptylammonium hydroxide or tetraheptylammonium chloride.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3536084A JPS60181096A (en) | 1984-02-28 | 1984-02-28 | Purification of bile acid |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3536084A JPS60181096A (en) | 1984-02-28 | 1984-02-28 | Purification of bile acid |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS60181096A true JPS60181096A (en) | 1985-09-14 |
JPH0364508B2 JPH0364508B2 (en) | 1991-10-07 |
Family
ID=12439713
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP3536084A Granted JPS60181096A (en) | 1984-02-28 | 1984-02-28 | Purification of bile acid |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS60181096A (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007069814A1 (en) * | 2005-12-12 | 2007-06-21 | Daewoong Pharmaceutical Co., Ltd. | Purification process for chenodeoxycholic acid |
WO2007078039A1 (en) * | 2005-12-30 | 2007-07-12 | Daewoong Pharmaceutical Co., Ltd. | Purification process for chenodeoxycholic acid |
CN103360454A (en) * | 2013-05-06 | 2013-10-23 | 广西大学 | Method for separating and purifying chenodeoxycholic acid from goose bile |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
TR201911039T4 (en) * | 2012-04-20 | 2019-08-21 | Ohr Pharmaceutical Inc | Aminosteroids for the treatment of a disease associated with Ptp1b. |
-
1984
- 1984-02-28 JP JP3536084A patent/JPS60181096A/en active Granted
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007069814A1 (en) * | 2005-12-12 | 2007-06-21 | Daewoong Pharmaceutical Co., Ltd. | Purification process for chenodeoxycholic acid |
WO2007078039A1 (en) * | 2005-12-30 | 2007-07-12 | Daewoong Pharmaceutical Co., Ltd. | Purification process for chenodeoxycholic acid |
CN103360454A (en) * | 2013-05-06 | 2013-10-23 | 广西大学 | Method for separating and purifying chenodeoxycholic acid from goose bile |
Also Published As
Publication number | Publication date |
---|---|
JPH0364508B2 (en) | 1991-10-07 |
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