CN103539829A - Method for extracting ursodesoxycholic acid - Google Patents
Method for extracting ursodesoxycholic acid Download PDFInfo
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- CN103539829A CN103539829A CN201310451578.4A CN201310451578A CN103539829A CN 103539829 A CN103539829 A CN 103539829A CN 201310451578 A CN201310451578 A CN 201310451578A CN 103539829 A CN103539829 A CN 103539829A
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- acid
- recrystallization
- ursodesoxycholic acid
- methyl esters
- chenodiol
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Abstract
The invention discloses a method for extracting ursodesoxycholic acid. The method for extracting ursodesoxycholic acid comprises the steps of esterification, extraction, recrystallization, hydrolysis, secondary recrystallization and the like. According to the method for extracting ursodesoxycholic acid, porcine bile is taken as an extraction raw material, after bilirubin is extracted from the porcine bile, filtrate after hyodeoxycholic acid and hyocholicacid are extracted is collected, and separation is carried out to obtain the ursodesoxycholic acid; porcine bile raw material is available, so that preparation cost of the ursodesoxycholic acid is reduced; acetic anhydride is selected as an esterifying reagent in an extraction process, ursodesoxycholic acid and recrystallization refining conditions are determined, sodium hydroxide is adopted as an alkali catalyst, and total yield of the ursodesoxycholic acid is more than 15% by adopting the technologic process.
Description
Technical field
The invention belongs to biological medicine technology field, be specifically related to a kind of extracting method of ursodesoxycholic acid.
Background technology
Bile is the product that human body or vertebrates liver cell secrete continuously, it is a kind of coloured alkaline liquid of viscosity with bitter taste, beyond its composition dewaters, mainly contain bile acide, cholate and bile pigment, also have in addition a small amount of cholesterol, lipid acid, Yelkin TTS, ionogen and protein etc.More common animal bile is generally bird gall, Fel Sus domestica, oxgall, Fel Ursi and Fel Serpentis etc. now.Bile acide is the main component of bile, account for 50%～70% of bile total amount, the physiological action of bile is mainly brought into play by bile acide exactly, when as medicine, that bile acide can be used for is analgesic, antispastic antibechic, regulate blood pressure, cardiac stimulant cholagogic, antibacterial anti-inflammatory and regulate immunity etc., and activity is strong, side effect is again little, cheap, is therefore subject to vast medication person's welcome.
In various animal biles, Fel Sus domestica is the most common and output is maximum, from ancient times, start to be just widely used in clinical, as by Fel Sus domestica treatment acute icteric infectious hepatitis, Whooping cough, acute enteritis and stomach ulcer etc., all can receive good therapeutic action.Main containing bile acids, bile pigment, Saliva Orthana, lipid and inorganics etc. in Fel Sus domestica, bile acide mainly comprises Hyodeoxycholic Acid, Iocholic acid, Chenodiol, 3 Alpha-hydroxies-6-oxygen-5 α-ursodeoxycholic acid and cholelith acid etc.Wherein Hyodeoxycholic Acid accounts for 40%, and Chenodiol accounts for 25%.
Ursodesoxycholic acid chemistry 3 α by name, 7 beta-dihydroxyl-5 β-ursodeoxycholic acids, molecular formula C
24h
49o
4, molecular weight is 392.58, is white or micro-yellow powder, is soluble in ethanol, glacial acetic acid, is slightly dissolved in chloroform, is dissolved in hardly ether and water.Ursodesoxycholic acid is first from Fel Ursi, to find nineteen twenty, and nineteen twenty-seven Shoda separation from Fel Ursi obtains crystal, and nineteen thirty-seven is determined its chemical structure.Natural ursodesoxycholic acid derives from Fel Ursi, and along with the demand of clinical application constantly increases, natural ursodesoxycholic acid can not be met the need of market far away.Kill bear and get courage and prohibited by country, therefore, by chemical synthesis process, obtain ursodesoxycholic acid and become more and more important.At first, it is raw material that cholic acid, Deoxycholic Acid etc. are take in people success, synthesized ursodesoxycholic acid, but this kind of method reactions steps is many, complex process, and productive rate is low, is difficult to carry out industrialization scale operation.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind ofly take Fel Sus domestica and is the method for the separated ursodesoxycholic acid of raw material, and the method extraction process is simple, and without specific installation, raw material sources are extensively easy to get, and environmental pollution is little, and product purity is high.
For solving the problems of the technologies described above, the technical scheme that Bian of the present invention gets is as follows:
An extracting method for ursodesoxycholic acid, comprises the steps:
(1) get and carried bilirubinic Fel Sus domestica tankage, through saponification, hydrolysis, decolouring and after being dried, obtain tan pig TOTAL BILE ACID crude product;
(2) get pig TOTAL BILE ACID crude product, after esterification, reagent obtains pig TOTAL BILE ACID ester powder with ethanol-extracted, recrystallization, then pig TOTAL BILE ACID ester is hydrolyzed under alkaline condition after, recrystallization obtains Hyodeoxycholic Acid white crystal for the second time;
(3) filtrate step (1) and (2) being obtained, washing lotion and recrystallization mother liquor merge, and decompression and solvent recovery obtains brown thick thing, adds diacetyl oxide esterifying reagent, carry out esterification and generate Iocholic acid ester under sodium acetate catalysis;
(4) under alkaline condition, with alkane, extract Iocholic acid methyl esters triacetate;
(5) mother liquor of above-mentioned recrystallization mother liquor and step (4) extraction Iocholic acid methyl esters triacetate is merged, removal of solvent under reduced pressure, obtains the thick thing of brown, after carrying with methyl alcohol, with ethyl alcohol recrystallization, obtain the white powder of Chenodiol methyl esters diacetate again;
(6) after Chenodiol methyl esters diacetate is hydrolyzed under alkaline condition, carry out recrystallization, under infrared lamp, be dried to after constant weight, make the white powder of Chenodiol.
As improvement of the present invention, in above-mentioned steps (5), the extraction time of methyl alcohol is 72～96 hours.
As improvement of the present invention, the recrystallization reagent of above-mentioned steps (5) is methyl alcohol or ethanol.
As improvement of the present invention, the alkali that above-mentioned steps (6) is used is NaOH, and the mol ratio of itself and pig Chenodiol methyl esters diacetate is 2.5:1.
As improvement of the present invention, above-mentioned steps (6) recrystallization reagent is ethyl acetate.
Compared with prior art, beneficial effect of the present invention is:
The present invention is usingd Fel Sus domestica as extracting raw material, and Fel Sus domestica extracts after bilirubin, collects extraction Hyodeoxycholic Acid and carries out separated with the filtrate after Iocholic acid and make ursodesoxycholic acid, and Fel Sus domestica raw material is extensively easy to get, and has reduced the preparation cost of ursodesoxycholic acid.In extraction process of the present invention, choosing diacetyl oxide is esterifying reagent, has determined the condition of ursodeoxycholic acid hydrolysis and recrystallizing and refining, determines and adopts sodium hydroxide as alkaline catalysts, and through this technical process, the total recovery of Iocholic acid reaches more than 15%.
Accompanying drawing explanation
Fig. 1 is process flow sheet of the present invention.
Embodiment
For ease of understanding the present invention, it is as follows that the present invention enumerates embodiment.Those skilled in the art should understand, described embodiment only, for helping to understand the present invention, should not be considered as concrete restriction of the present invention.
In the following example, the same meaning that all specialties of using and scientific words and those skilled in the art are familiar.
As shown in Figure 1, a kind of extracting method of ursodesoxycholic acid, comprises the steps:
(1) get and carried bilirubinic Fel Sus domestica tankage, through saponification, hydrolysis, decolouring and after being dried, obtain tan pig TOTAL BILE ACID crude product;
(2) get pig TOTAL BILE ACID crude product, after esterification, reagent obtains pig TOTAL BILE ACID ester powder with ethanol-extracted, recrystallization, then pig TOTAL BILE ACID ester is hydrolyzed under alkaline condition after, recrystallization obtains Hyodeoxycholic Acid white crystal for the second time;
(3) filtrate step (1) and (2) being obtained, washing lotion and recrystallization mother liquor merge, and decompression and solvent recovery obtains brown thick thing, adds diacetyl oxide esterifying reagent, carry out esterification and generate Iocholic acid ester under sodium acetate catalysis;
(4) under alkaline condition, with alkane, extract Iocholic acid methyl esters triacetate;
(5) mother liquor of above-mentioned recrystallization mother liquor and step (4) extraction Iocholic acid methyl esters triacetate is merged, removal of solvent under reduced pressure, obtains the thick thing of brown, after carrying with methyl alcohol, with ethyl alcohol recrystallization, obtain the white powder of Chenodiol methyl esters diacetate again;
(6) after Chenodiol methyl esters diacetate is hydrolyzed under alkaline condition, carry out recrystallization, under infrared lamp, be dried to after constant weight, make the white powder of Chenodiol.
Embodiment 1
(1) get 250ml round-bottomed flask, add pig TOTAL BILE ACID crude product 10g, anhydrous methanol 50ml, 98% vitriol oil 0.7ml, stirring reaction 24h under normal temperature, complete through TLC detection reaction, without the residual rear stopped reaction of raw material, in system, add anhydrous Na HCO
3solid adjusts pH to neutral, elimination inorganic salt, filter cake methanol wash, merging filtrate and washing lotion, reclaim under reduced pressure methyl alcohol, obtain brown dope, add water 50ml to dissolve wherein remaining inorganic salt, then extract organism (50ml with ether gradation, 30ml, 20ml), merge organic phase, respectively with saturated NaHCO
3solution and saturated common salt water washing 1 time, anhydrous Na
2sO
4dry 2h, filters, and removes ether under reduced pressure, obtain brown thick thing 11.4g, in flask, add benzene 30ml, at 80 ℃ of oil bath pans, after dissolve complete, stop heating, after naturally cooling to room temperature, put into the about 16h of refrigerator, separate out solid, filter, filter cake is with benzene washing 2～3 times, obtain off-white color solid 4.9g, then carry out recrystallization with benzene, the white powder obtaining is dried to constant weight under infrared lamp, for 4.5g, i.e. pig TOTAL BILE ACID methyl esters;
(2) get 100ml round-bottomed flask, add successively Hyodeoxycholic Acid methyl esters (4.5g), NaOH solid (1.1g), ethanol 30ml, water 1ml, stirring and refluxing reaction 20h at 80 ℃, after completion of the reaction, decompression and solvent recovery, to dry, obtains faint yellow solid in TLC monitoring, add water 30ml, under vigorous stirring, add 10% dilute hydrochloric acid and adjust pH to 1.5, separate out a large amount of solids, continue to stir at normal temperatures 0.5h, filter, 10% dilute hydrochloric acid washing 3 times for filter cake, washes 3 times, obtains block white solid, drying under reduced pressure 8h at 50 ℃ is 4.5g.With re-crystallizing in ethyl acetate, after the white crystal obtaining is dried to constant weight under infrared lamp, be 4g, mp is 200～201 ℃, i.e. Hyodeoxycholic Acid.
(3) filtrate upper step being obtained, washing lotion and recrystallization mother liquor merge, and reclaim under reduced pressure benzene, to the greatest extent, obtains brown thick thing 6.7g.Add diacetyl oxide 54ml, sodium acetate 8g, back flow reaction 5h at 110 ℃, after TLC monitoring has been reacted, remove the diacetyl oxide of approximately 2/3 volume under reduced pressure, after system is down to room temperature naturally, add water 50ml to dissolve inorganic salt wherein, with ether gradation, extract organism (50ml, 30ml, 20ml), merge organic phase, first neutral to approaching by 1% NaOH solution washing organic phase, use again saturated NaHCO3 solution and saturated common salt water washing, anhydrous Na
2sO
4dry 2h, filters, and ether is removed in decompression, obtain brown thick thing 7.9g, in flask, add normal hexane 20ml, system is warming up to after 65 ℃ of stirrings are dissolved it completely and stops heating, naturally be down to room temperature, at room temperature stir 2h, filter, normal hexane washing 3 times for filter cake, after dry, obtain brown ceramic powder 1.7g, successively with methyl alcohol and benzene-sherwood oil system recrystallization, after being dried, obtain white powder 0.8g, mp184～186 ℃, i.e. Iocholic acid methyl esters triacetate.
(4) get 25ml round-bottomed flask, add successively Iocholic acid methyl esters triacetate (0.8g), NaOH solid (0.35g), ethanol 10ml, water 0.4ml, back flow reaction 20h at 80 ℃, TLC monitoring after completion of the reaction, decompression and solvent recovery, obtain pale solid, add water 10ml, under vigorous stirring, with 10% dilute hydrochloric acid, adjust PH to 1.52, separate out solid, continue under normal temperature and stir 0.5h, filter, filter cake respectively washs 3 times with 10% dilute hydrochloric acid and water successively, obtain white solid, at 50 ℃, after drying under reduced pressure, be 0.6g, with re-crystallizing in ethyl acetate, under infrared lamp, be dried to constant weight, obtain white powder 0.52g, mp188～189 ℃, be Iocholic acid.
(5) mother liquor and the recrystallization mother liquor that have extracted Iocholic acid methyl esters triacetate are merged, removal of solvent under reduced pressure, obtain the thick thing 6.3g of brown, now wherein main component is Chenodiol methyl esters diacetate, in flask, add methyl alcohol 50ml, be warming up to 65 ℃ and stop heating after dope all dissolves, system is down to after room temperature standing 72h in refrigerator naturally, separate out faint yellow solid, filter, filter cake washs by ice cold methanol, after dry, obtain pale yellow powder 2.4g, with ethyl alcohol recrystallization, solid is dried to constant weight under infrared lamp, obtain white powder 2.1g, mp111～113 ℃, be Chenodiol methyl esters diacetate.
(6) get 50ml round-bottomed flask, add successively Chenodiol methyl esters diacetate (2.1g), NaOH solid (0.9g), ethanol 20ml, water 1ml, back flow reaction 20h at 80 ℃, TLC monitoring after completion of the reaction, decompression and solvent recovery, obtain pale solid, add water 20ml, under vigorous stirring, with 10% dilute hydrochloric acid, adjust PH to 1.5, separate out solid, continue under normal temperature and stir 0.5h, filter, filter cake respectively washs 3 times with 10% dilute hydrochloric acid and water successively, obtain white solid, at 50 ℃, after drying under reduced pressure, be 1.6g, with re-crystallizing in ethyl acetate, after being dried to constant weight under infrared lamp, obtain white powder 1.6g, mp141～144 ℃, be Chenodiol, total recovery is 16%.
Embodiment 2
Compare with embodiment 1, the (1), the (2nd) (3), (4 steps are identical, and (5) and (6) step is as follows:
(5) mother liquor and the recrystallization mother liquor that have extracted Iocholic acid methyl esters triacetate are merged, removal of solvent under reduced pressure, obtain the thick thing 6.3g of brown, now wherein main component is Chenodiol methyl esters diacetate, in flask, add methyl alcohol 50ml, be warming up to 65 ℃ and stop heating after dope all dissolves, system is down to after room temperature standing 96h in refrigerator naturally, separate out faint yellow solid, filter, filter cake washs by ice cold methanol, after dry, obtain pale yellow powder 2.4g, with recrystallizing methanol, solid is dried to constant weight under infrared lamp, obtain white powder 2.1g, mp111～113 ℃, be Chenodiol methyl esters diacetate.
(6) get 50ml round-bottomed flask, add successively Chenodiol methyl esters diacetate (2.1g), NaOH solid (0.9g), ethanol 20ml, water 1ml, back flow reaction 20h at 80 ℃, TLC monitoring after completion of the reaction, decompression and solvent recovery, obtain pale solid, add water 20ml, under vigorous stirring, with 10% dilute hydrochloric acid, adjust PH to 1.5, separate out solid, continue under normal temperature and stir 0.5h, filter, filter cake respectively washs 3 times with 10% dilute hydrochloric acid and water successively, obtain white solid, at 50 ℃, after drying under reduced pressure, be 1.6g, with re-crystallizing in ethyl acetate, after being dried to constant weight under infrared lamp, obtain white powder 1.5g, mp141～144 ℃, be Chenodiol, total recovery is 15%.
Applicant's statement, person of ordinary skill in the field is on the basis of above-described embodiment, by the concrete content point value of above-described embodiment component, combined with the technical scheme of summary of the invention part, thereby the new numerical range producing, also be one of record scope of the present invention, the application, for making specification sheets simple and clear, is no longer enumerated these numerical ranges.
Claims (5)
1. an extracting method for ursodesoxycholic acid, is characterized in that, comprises the steps:
(1) get and carried bilirubinic Fel Sus domestica tankage, through saponification, hydrolysis, decolouring and after being dried, obtain tan pig TOTAL BILE ACID crude product;
(2) get pig TOTAL BILE ACID crude product, after esterification, reagent obtains pig TOTAL BILE ACID ester powder with ethanol-extracted, recrystallization, then pig TOTAL BILE ACID ester is hydrolyzed under alkaline condition after, recrystallization obtains Hyodeoxycholic Acid white crystal for the second time;
(3) filtrate step (1) and (2) being obtained, washing lotion and recrystallization mother liquor merge, and decompression and solvent recovery obtains brown thick thing, adds diacetyl oxide esterifying reagent, carry out esterification and generate Iocholic acid ester under sodium acetate catalysis;
(4) under alkaline condition, with alkane, extract Iocholic acid methyl esters triacetate;
(5) mother liquor of above-mentioned recrystallization mother liquor and step (4) extraction Iocholic acid methyl esters triacetate is merged, removal of solvent under reduced pressure, obtains the thick thing of brown, after carrying, then with alcohol recrystallization, obtains the white powder of Chenodiol methyl esters diacetate with methyl alcohol;
(6) after Chenodiol methyl esters diacetate is hydrolyzed under alkaline condition, carry out recrystallization, under infrared lamp, be dried to after constant weight, make the white powder of Chenodiol.
2. the extracting method of a kind of ursodesoxycholic acid as claimed in claim 1, is characterized in that: in described step (5), the extraction time of methyl alcohol is 72～96 hours.
3. the extracting method of a kind of ursodesoxycholic acid as claimed in claim 1, is characterized in that: the recrystallization reagent of described step (5) is methyl alcohol or ethanol.
4. the extracting method of a kind of ursodesoxycholic acid as claimed in claim 1, is characterized in that: the alkali that described step (6) is used is NaOH, and the mol ratio of itself and pig Chenodiol methyl esters diacetate is 2.5:1.
5. the extracting method of a kind of ursodesoxycholic acid as claimed in claim 1, is characterized in that: described step (6) recrystallization reagent is ethyl acetate.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108522859A (en) * | 2018-07-09 | 2018-09-14 | 山东龙昌动物保健品有限公司 | Application of the bile acid in preparing the feed addictive for Broiler chicks |
| CN109722463A (en) * | 2019-03-18 | 2019-05-07 | 常德云港生物科技有限公司 | A method of hyocholic acid is synthesized by ursodesoxycholic acid by half enzyme process |
| CN110790805A (en) * | 2019-11-14 | 2020-02-14 | 湖南九典制药股份有限公司 | Method for extracting chenodeoxycholic acid from pig bile paste |
-
2013
- 2013-09-27 CN CN201310451578.4A patent/CN103539829A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 丁新腾等: "《从猪胆汁提取鹅去氧胆酸及猪胆酸》", 《医药工业》 * |
| 邓启华等: "《猪胆汁中三种主要胆汁酸的提取分离》", 《中国生化药物杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108522859A (en) * | 2018-07-09 | 2018-09-14 | 山东龙昌动物保健品有限公司 | Application of the bile acid in preparing the feed addictive for Broiler chicks |
| CN109722463A (en) * | 2019-03-18 | 2019-05-07 | 常德云港生物科技有限公司 | A method of hyocholic acid is synthesized by ursodesoxycholic acid by half enzyme process |
| CN110790805A (en) * | 2019-11-14 | 2020-02-14 | 湖南九典制药股份有限公司 | Method for extracting chenodeoxycholic acid from pig bile paste |
| CN110790805B (en) * | 2019-11-14 | 2022-08-30 | 湖南九典制药股份有限公司 | Method for extracting chenodeoxycholic acid from pig bile paste |
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Application publication date: 20140129 |