CN110845564A - Method for extracting chenodeoxycholic acid from chicken gall paste - Google Patents
Method for extracting chenodeoxycholic acid from chicken gall paste Download PDFInfo
- Publication number
- CN110845564A CN110845564A CN201911091094.7A CN201911091094A CN110845564A CN 110845564 A CN110845564 A CN 110845564A CN 201911091094 A CN201911091094 A CN 201911091094A CN 110845564 A CN110845564 A CN 110845564A
- Authority
- CN
- China
- Prior art keywords
- acid
- silica gel
- chenodeoxycholic acid
- chicken
- paste
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
- C07J9/005—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Steroid Compounds (AREA)
Abstract
The application discloses a method for extracting chenodeoxycholic acid from chicken bile paste, which comprises the steps of saponification, acid-adjusting extraction, methyl esterification, acetylation and the like, and achieves the purpose of separating cholic acid and chenodeoxycholic acid from the chicken bile paste at one time. The two products obtained by the method have high purity, the reagents are cheap and easy to obtain, most of solvents can be recycled, and the generated wastewater is less.
Description
Technical Field
The invention belongs to the field of separation and purification, and particularly relates to a method for extracting chenodeoxycholic acid from chicken gall paste.
Background
Chenodeoxycholic Acid (CDCA) with chemical name of 3 α,7 α -dihydroxy-5 β -cholanic acid (CAS number: 474-25-9) and molecular formula of C24H40O4Molecular weight is 392.57, and the structural formula is shown as follows:
。
chenodeoxycholic acid is a substance mainly extracted from bile, is mainly used for reducing the saturation of cholesterol in the bile, is mainly used for treating gallstones clinically, is also a main raw material for synthesizing ursodeoxycholic acid (UDCA) at present, is formally developed for treating cholesterol stones in 1972 by the UK, and is applied as a first medicament for correcting saturated bile and dissolving gallstones.
Chinese patent CN 201810179875.0 discloses a method for synthesizing ursodeoxycholic acid from chenodeoxycholic acid and cholic acid in chicken gallbladder, which comprises the steps of extraction, salt-forming crystallization, oxidation, reduction and Huang Minglong reaction. The method of the invention reduces the steps of extracting chenodeoxycholic acid from chicken bile acid, purifying and then synthesizing ursodesoxycholic acid in the prior art, greatly shortens the process links and saves the cost. The waste cholic acid discarded by the prior art is used as a widened raw material source and reacts with the chenodeoxycholic acid in the chicken bile at the same time to synthesize the ursodesoxycholic acid crude product, so that waste is changed into valuable, the utilization rate of the chenodeoxycholic acid is greatly improved, limited biological resources are developed and utilized to the maximum extent, and the technical problems of complex process, high cost, low yield and more waste in the prior art are solved.
The chicken bile paste is a waste of chicken product processing enterprises, is large in quantity, is easy to obtain raw materials, and mainly comprises cholic acid and chenodeoxycholic acid. The main difficulty of the technology for extracting chenodeoxycholic acid from chicken bile paste is the separation of chenodeoxycholic acid (CDCA) and Cholic Acid (CA), wherein the structure of the cholic acid is shown as the following formula:
as shown by the structural comparison, Cholic Acid (CA) has one more hydroxyl group than chenodeoxycholic acid (CDCA), and the structural similarity makes the separation of two substances difficult.
Chenodeoxycholic acid is the main component of fowl bile such as chicken, duck and goose, and can be directly extracted from fowl bile. The chenodeoxycholic acid products in the current market are basically extracted from chicken gall, but are limited by the technology, the existing extraction methods mainly comprise a barium salt method and a calcium salt precipitation method, a crystallization extraction method and an ion exchange resin method, the barium salt can bring serious environmental problems, when the calcium salt is used, a great deal of water is used, water resources are greatly wasted, the crystallization extraction method needs to use a great amount of organic solvent, mother liquor needs to be separated and crystallized repeatedly, the production period is long, large-scale production cannot be realized, the extraction yield of the methods is low, and the purity of the obtained products is generally lower than 90%.
Therefore, under the circumstances that the prior art has the above problems, it is very significant to develop an extraction technology which can effectively extract cholic acid and chenodeoxycholic acid from chicken bile paste at one time and has the advantages of good effect, high efficiency, low cost, high purity and the like.
Disclosure of Invention
The main purpose of the application is to provide a method for extracting cholic acid and chenodeoxycholic acid from chicken bile paste.
The technical scheme of the method is based on the purpose of extracting chenodeoxycholic acid from the chicken gall paste, and is formed by further optimizing and improving the prior art. In order to achieve the above objects and solve the problems in the prior art, the present application adopts the following technical solutions:
a method for extracting chenodeoxycholic acid from chicken bile paste comprises the steps of saponification, acid-adjusting extraction, methyl esterification, acetylation, silica gel filtration, reflux hydrolysis and the like, wherein the filtration mode of the silica gel filtration step is that silica gel is paved in a filter funnel for filtration.
Preferably, the thickness of the silica gel is 2-5 cm or 8-12 cm, and the silica gel is applied to a pilot plant test and a pilot plant test respectively.
Preferably, the method also comprises a mother liquor recovery step, wherein the mother liquor recovery step comprises the following steps: the crystallized mother liquor is concentrated to remove the alkane solvent and then is reused in the step of acid regulation and extraction before the next batch of methyl esterification.
Through multiple experiments, the experimenter finds out a better experimental scheme for extracting cholic acid and chenodeoxycholic acid from the chicken gall paste at one time, and the experimental scheme is as follows:
a method for extracting chenodeoxycholic acid from chicken bile paste comprises the following steps:
(1) saponification: adding sodium hydroxide and water into the chicken bile paste, heating and refluxing for saponification to obtain a reaction solution, wherein the original chicken bile paste contains a large amount of amino acids, and the saponification is carried out to dissociate cholic acid and chenodeoxycholic acid from a bound state.
(2) Extraction and esterification: adding an ester solvent into the reaction liquid obtained in the step (1), adding ethyl acetate, adjusting acidity for extraction, concentrating the extract until the reaction system becomes oily, adding methanol and concentrated sulfuric acid for reflux stirring, esterifying, adding solid sodium bicarbonate for adjusting pH after esterification, filtering to obtain a bile acid methyl ester methanol solution, concentrating to obtain oily matter, extracting chenodeoxycholic acid and cholic acid from an aqueous phase to an organic phase through extraction, further eliminating impurities, and changing carboxyl groups in the cholic acid and the chenodeoxycholic acid into methyl ester groups through esterification to change the polarities of the cholic acid and the chenodeoxycholic acid.
(3) Acetylation: adding an acetylation reagent into the oily substance obtained in the step (2) to perform heating reflux reaction, cooling to room temperature after TLC detection reaction is finished, filtering, and concentrating the filtrate to obtain an oily substance, wherein hydroxyl groups in cholic acid and chenodeoxycholic acid are acetylated in the step to further change the polarity of the cholic acid and chenodeoxycholic acid.
(4) Silica gel filtration: adding an alkane solvent into the oily matter obtained in the step (3), heating to completely dissolve the oily matter obtained in the step (3), adding silica gel powder, cooling, spreading a layer of silica gel in a filter funnel for filtering, washing with the alkane solvent once to obtain a filtrate, wherein a filter cake is triacetyl cholic acid methyl ester, hydrolyzing the triacetyl cholic acid methyl ester recovered from the filter cake to obtain cholic acid, wherein the polarity difference between the triacetyl cholic acid methyl ester and diacetyl chenodeoxycholic acid methyl ester is utilized, the difference of silica gel adsorption is utilized, the triacetyl cholic acid methyl ester with larger polarity is adsorbed, and the diacetyl chenodeoxycholic acid is eluted.
(5) Refluxing and hydrolyzing: concentrating the filtrate obtained in the step (4) to recover the solvent, adding a sodium hydroxide aqueous solution for heating reflux hydrolysis, cooling to adjust acidity, adding ethyl acetate for extraction, continuously adding acid to adjust pH, heating, stirring for standing for layering, removing a water layer, cooling again for crystallization, and dropwise adding an alkane solvent to expel crystals to obtain chenodeoxycholic acid.
(6) Mother liquor recovery: and concentrating the crystallization mother liquor to remove the alkane solvent, and then, mechanically applying the crystallization mother liquor to the next batch of extraction liquid to perform an esterification step.
Preferably, the temperature for heating reflux to carry out saponification in the step (1) is 95-105 ℃, and the saponification duration is 8-15 hours.
Preferably, the acetylation reagent in the step (3) is one selected from acetic anhydride, sodium acetate combination or acetyl chloride and triethylamine combination, the temperature is raised to 120-140 ℃, and the reflux time is 3-4 hours.
Preferably, the specification of the silica gel in the step (4) is selected from one or more of 100-200 meshes, 200-300 meshes, 300-400 meshes and 400-500 meshes. The thickness of the silica gel depends on the weight of the material, and generally, the silica gel is 2 to 5cm thick in a laboratory pilot scale and 8 to 12cm thick in a factory pilot scale.
Preferably, the temperature for dissolving the oily substance by heating in the step (4) is 60-70 ℃.
Preferably, the temperature of the silica gel powder added in the step (4) is 20-40 ℃.
Preferably, the mass of the silica gel powder added in the step (4) is 0.1-1 time of that of the chicken gall paste.
Preferably, the heating temperature for dissolving the oily substance by adding the alkane solvent in the step (4) is 40-70 ℃.
Preferably, the temperature of the heating reflux in the step (5) is 80-120 ℃, and the reflux time is 8-15 lab tests.
Preferably, the temperature for cooling in the step (5) is 40-50 ℃, the pH value of acidity is adjusted to 7-8, the pH value is continuously adjusted to 2-4 by adding acid, the temperature for heating and stirring is 40-60 ℃, and the temperature for cooling and crystallizing is-15-5 ℃.
Preferably, the alkane solvent in step (4) and step (5) is one or more of hexane, heptane, dichloromethane, cyclohexane and petroleum ether.
The application also comprises cholic acid and chenodeoxycholic acid prepared by the method.
The application also includes the use of the method in the purification of chenodeoxycholic acid.
The main difficulty of the technology for extracting the chenodeoxycholic acid from the chicken bile paste is the separation of cholic acid and chenodeoxycholic acid. The structure of cholic acid is very similar to that of chenodeoxycholic acid, and the inventor surprisingly finds that a methyl ester acetylation product triacetyl cholic acid methyl ester of cholic acid and a methyl ester acetylation product diacetyl chenodeoxycholic acid methyl ester of chenodeoxycholic acid can be well separated when the methyl ester acetylation product triacetyl cholic acid methyl ester and the methyl ester acetylation product diacetyl chenodeoxycholic acid methyl ester are filtered by silica gel, triacetyl cholic acid methyl ester with larger polarity can be adsorbed by the silica gel, diacetyl chenodeoxycholic acid methyl ester with smaller polarity is eluted, and therefore. And hydrolyzing the triacetyl cholic acid methyl ester adsorbed by the silica gel to obtain the cholic acid. The rest filtrate is diacetyl chenodeoxycholic acid methyl ester, and chenodeoxycholic acid can be obtained by reflux hydrolysis, extraction, cooling and crystallization. Therefore, the whole scheme can separate and extract two substances of cholic acid and chenodeoxycholic acid at one time, not only can effectively obtain the target product chenodeoxycholic acid, but also can use the separated cholic acid for other purposes, does not cause material waste, and has excellent separation effect through silica gel.
The extraction method of our prescription also solves a plurality of technical problems of the chenodeoxycholic acid in the past production process. For example, in the actual production process of chenodeoxycholic acid, the precipitation of the oily substance in the step (4) is a problem influencing the experimental result, and in the past production process, if the oily substance in the step (4) is directly dissolved and heated by the alkane solvent and then is directly cooled, in the process, the oily substance is very easily separated out again, so that the material of the chenodeoxycholic acid is lost and the subsequent silica gel filtration is influenced. Thereby my side has improved technical scheme, has added the silica gel powder for oily thing can not appear at the in-process of cooling, has solved the problem that oily thing appears easily among the cooling process. The extraction esterification and acetylation in the steps of the method are all group modification on products to be separated, and related polarity and physicochemical properties are changed, so that the final silica gel filtration step is facilitated.
Due to the adoption of the technical scheme, the beneficial effects of the application are as follows:
1. the separation of the two components is realized by utilizing the difference of the adsorption degrees of triacetyl cholic acid methyl ester and diacetyl chenodeoxycholic acid methyl ester in silica gel, the separation effect is very good, the content of the cholic acid in the obtained chenodeoxycholic acid crude product is lower than 5%, and the content of the cholic acid in the chenodeoxycholic acid crude product is lower than 1% after the chenodeoxycholic acid crude product is refined.
2. The cholic acid separated in the chenodeoxycholic acid extraction process has high purity and can be used for other purposes.
3. The reagent used in the process of extracting chenodeoxycholic acid is cheap and easy to obtain, most of the solvent can be recycled, and the generated wastewater is less.
4. The application of the method can improve the extraction yield of the chenodeoxycholic acid by extracting and recycling the mother liquor in the extraction process of the chenodeoxycholic acid.
Drawings
FIG. 1 is a flow chart of my party solution;
FIG. 2 is a detection spectrum of a finished product prepared in example 1 of my party;
FIG. 3 is a detection profile of the filtrate after saponification of my example 1;
FIG. 4 is a detection spectrum of the filtrate after silica gel filtration of my example 1;
FIG. 5 is a detection spectrum of the finished product prepared in example 2 of my party;
FIG. 6 is a detection spectrum of the finished product prepared in my example 3.
Detailed Description
The present application is described in further detail below with reference to specific embodiments and the attached drawings. The following examples are intended to be illustrative of the present application only and should not be construed as limiting the present application.
Example 1
A method for extracting chenodeoxycholic acid from chicken bile paste is an example of a laboratory experiment and comprises the following steps:
(1) saponification: 1.8 kg of chicken gall cream (the content of CDCA is 24.5%) is put into a 30L reaction kettle, 6.0 kg of water and 0.54 kg of sodium hydroxide are added, the mixture is heated to reflux, and the reflux and heat preservation are carried out for 8 hours to be cooled to room temperature.
(2) Extraction and esterification: dropwise adding hydrochloric acid into the reaction liquid obtained in the previous step to adjust the pH = 7-8, adding 7.2L of ethyl acetate, continuously dropwise adding hydrochloric acid to adjust the pH = 2-3, heating to 40-50 ℃, fully stirring for 1 hour, standing for layering, and removing a water layer to obtain an ethyl acetate solution. Concentrating the ethyl acetate solution to an oily substance, adding 4L of methanol, dripping 0.07kg of concentrated sulfuric acid, heating and refluxing for 3h, adding a sodium bicarbonate solid after TLC detection reaction is completed, adjusting the pH to be 7-8, filtering, washing a filter cake with 0.4L of methanol, and concentrating the filtrate to the oily substance.
(3) Acetylation: adding 2L of acetic anhydride and 0.1kg of sodium acetate into the oily substance obtained in the step (3), heating to reflux, detecting by TLC after 3 hours until the reaction is complete, cooling to room temperature, filtering, and concentrating the filtrate to obtain the oily substance.
(4) Silica gel filtration: adding 8L of heptane into the oily matter obtained in the step (4), heating, refluxing, dissolving, adding 300g of silica gel powder to prevent the oily matter from being separated out again in the cooling process, cooling to 20 ℃, filtering through silica gel (the silica gel is paved in a filtering funnel and has the thickness of about 2-5 cm), leaching with 4L of heptane to obtain filtrate and filter cake, and adding water and sodium hydroxide into the triacetyl cholic acid methyl ester recovered from the filter cake to hydrolyze to obtain cholic acid.
(5) Refluxing and hydrolyzing: concentrating the filtrate obtained in the step (5) to an oily substance, adding 0.2kg of sodium hydroxide and 2kg of water, heating and refluxing for 12 hours, finishing TLC detection reaction, cooling to 50 ℃, adjusting the pH to be 6-7, adding 2L of ethyl acetate, continuously dropwise adding hydrochloric acid to adjust the pH to be 2-3, heating to 50-60 ℃, fully stirring for 1 hour, standing for layering, discarding a water layer, cooling to 0-5 ℃ to separate out a solid, dropwise adding 4L of dichloromethane, stirring for 2 hours, filtering to obtain 0.6 kg of a wet chenodeoxycholic acid product, and drying to obtain 0.35 kg of a finished chenodeoxycholic acid product.
(6) Mother liquor recovery: and concentrating the crystallization mother liquor to remove the alkane solvent, and then, mechanically applying the crystallization mother liquor to the next batch of extraction liquid to perform an esterification step.
Inspecting the finished productThe test results are shown in the following table:
| variety of (IV) C | Chenodeoxycholic acid | Cholic acid |
| Purity of | 98.68% | 0.77% |
The detection spectrum is shown in fig. 2, and therefore, the purity of the chenodeoxycholic acid prepared by the method is very high, and the chenodeoxycholic acid can reach the level of more than 97% after refining. The content of cholic acid and other impurities is also very low, and is below 1 percent, and the content of the impurities is well controlled.
The method also takes key nodes in the preparation process for detection, and the detection results are shown in the following graphs in fig. 3-4 respectively after saponification and silica gel filtration hydrolysis are finished:
after saponification
| Variety of (IV) C | Chenodeoxycholic acid | Cholic acid |
| Purity of | 72.76% | 20.27% |
After silica gel filtration and hydrolysis
| Variety of (IV) C | Chenodeoxycholic acid | Cholic acid |
| Purity of | 92.00% | 1.52% |
From the above table, we know that the effect of removing cholic acid is significant.
Example 2
A method for extracting chenodeoxycholic acid from chicken bile paste is an example of a laboratory experiment and comprises the following steps:
(1) saponification: 1.8 kg of chicken gall cream (the content of CDCA is 24.5%) is put into a 30L reaction kettle, 6.0 kg of water and 0.54 kg of sodium hydroxide are added, the mixture is heated to reflux, and the reflux and heat preservation are carried out for 8 hours to be cooled to room temperature.
(2) Extraction and esterification: dropwise adding hydrochloric acid into the reaction liquid obtained in the previous step to adjust the pH = 7-8, adding 7.2L of butyl acetate, continuously dropwise adding hydrochloric acid to adjust the pH = 2-3, heating to 40-50 ℃, fully stirring for 1 hour, standing for layering, and removing a water layer to obtain a butyl acetate solution. Concentrating a butyl acetate solution to an oily substance, adding 4L of methanol, dripping 0.07kg of concentrated sulfuric acid, heating and refluxing for 3h, adding a sodium bicarbonate solid after TLC detection reaction is completed, adjusting the pH to be 7-8, filtering, washing a filter cake with 0.4L of methanol, and concentrating a filtrate to an oily substance.
(3) Acetylation: adding 2L of acetic anhydride and 0.1kg of sodium acetate into the oily substance obtained in the step (3), heating to reflux, detecting by TLC after 3 hours until the reaction is complete, cooling to room temperature, filtering, and concentrating the filtrate to obtain the oily substance.
(4) Silica gel filtration: adding 8L of n-hexane into the oily matter obtained in the step (4), heating, refluxing, dissolving and clearing, adding 300g of silica gel powder to prevent the oily matter from being separated out again in the cooling process, cooling to 20 ℃, filtering through silica gel (the silica gel with the thickness of about 3-5 cm is paved in a filter funnel), leaching with 4L of n-hexane to obtain filtrate and filter cake, and adding water and sodium hydroxide into the triacetyl cholic acid methyl ester recovered from the filter cake to hydrolyze to obtain cholic acid.
(5) Refluxing and hydrolyzing: concentrating the filtrate obtained in the step (5) to an oily substance, adding 0.2kg of sodium hydroxide and 2kg of water, heating and refluxing for 12 hours, finishing TLC detection reaction, cooling to 50 ℃, adjusting the pH to be 6-7, adding 2L of butyl acetate, continuously dropwise adding hydrochloric acid to adjust the pH to be 2-3, heating to 50-60 ℃, fully stirring for 1 hour, standing for layering, discarding a water layer, cooling to 0-5 ℃ to separate out a solid, dropwise adding 4L of dichloromethane, stirring for 2 hours, filtering to obtain 0.65 kg of a wet chenodeoxycholic acid product, and drying to obtain 0.37 kg of a finished chenodeoxycholic acid product.
(6) Mother liquor recovery: and concentrating the crystallization mother liquor to remove the alkane solvent, and then, mechanically applying the crystallization mother liquor to the next batch of extraction liquid to perform an esterification step.
The finished product was submitted for inspection and the results are shown in the following table:
| variety of (IV) C | Chenodeoxycholic acid | Cholic acid |
| Purity of | 98.59% | 0.47% |
The detection pattern is shown in fig. 5, and therefore, the purity of the chenodeoxycholic acid prepared by the method is very high, and the chenodeoxycholic acid can reach the level of more than 97% after refining. The content of cholic acid and other impurities is also very low, and is below 1 percent, and the content of the impurities is well controlled.
Example 3
A method for extracting chenodeoxycholic acid from chicken bile paste is an example for carrying out a pilot test, and comprises the following steps:
(1) saponification: 90 kg of chicken gall cream (the content of CDCA is 24.5%) is put into a 1000L reaction kettle, 300 kg of water and 27 kg of sodium hydroxide are added, the mixture is heated to reflux, and the reflux and heat preservation are carried out for 8 hours to be cooled to room temperature.
(2) Extraction and esterification: and (3) dropwise adding hydrochloric acid into the reaction liquid obtained in the previous step to adjust the pH = 7-8, adding 360L of isopropyl acetate, continuously dropwise adding hydrochloric acid to adjust the pH = 2-3, heating to 40-50 ℃, fully stirring for 1 hour, standing for layering, and removing a water layer to obtain an isopropyl acetate solution. Concentrating an isopropyl acetate solution to an oily substance, adding 200L of methanol, dripping 3.5kg of concentrated sulfuric acid, heating and refluxing for 3h, adding a sodium bicarbonate solid after TLC detection reaction is completed to adjust the pH to be 7-8, filtering, washing a filter cake with 20L of methanol, and concentrating a filtrate to an oily substance.
(3) Acetylation: and (4) adding 100L of acetic anhydride and 5kg of sodium acetate into the oily substance obtained in the step (3), heating to reflux, detecting by TLC after 3 hours until the reaction is complete, cooling to room temperature, filtering, and concentrating the filtrate to obtain the oily substance.
(4) Silica gel filtration: adding 400L of heptane into the oily matter obtained in the step (4), heating, refluxing, dissolving, adding 15kg of silica gel powder to prevent the oily matter from being separated out again in the cooling process, cooling to 20 ℃, filtering through silica gel (the silica gel is paved in a filtering funnel with the thickness of about 3-5 cm), leaching with 200L of heptane to obtain filtrate and filter cake, and adding water and sodium hydroxide into the triacetyl cholic acid methyl ester recovered from the filter cake to hydrolyze to obtain the cholic acid.
(5) Refluxing and hydrolyzing: concentrating the filtrate obtained in the step (5) to an oily substance, adding 0.2kg of sodium hydroxide and 2kg of water, heating and refluxing for 12 hours, finishing TLC detection reaction, cooling to 50 ℃, adjusting the pH to be 6-7, adding 2L of isopropyl acetate, continuously dropwise adding hydrochloric acid to adjust the pH to be 2-3, heating to 50-60 ℃, fully stirring for 1 hour, standing for layering, discarding a water layer, cooling to 0-5 ℃, precipitating a solid, dropwise adding 4L of dichloromethane, stirring for 2 hours, filtering to obtain 32 kg of wet chenodeoxycholic acid product, and drying to obtain 18.3 kg of finished chenodeoxycholic acid product.
(6) Mother liquor recovery: and concentrating the crystallization mother liquor to remove the alkane solvent, and then, mechanically applying the crystallization mother liquor to the next batch of extraction liquid to perform an esterification step.
The finished product was submitted for inspection and the results are shown in the following table:
| variety of (IV) C | Chenodeoxycholic acid | Cholic acid |
| Purity of | 98.33% | 0.73% |
The detection pattern is shown in fig. 6, and therefore, the purity of the chenodeoxycholic acid prepared by the method is very high, and the chenodeoxycholic acid can reach the level of more than 97% after refining. The content of cholic acid and other impurities is also very low, and is below 1 percent, and the content of the impurities is well controlled.
The foregoing is a more detailed description of the present application in connection with specific embodiments thereof, and it is not intended that the present application be limited to the specific embodiments thereof. For those skilled in the art to which the present application pertains, several simple deductions or substitutions may be made without departing from the concept of the present application, and all should be considered as belonging to the protection scope of the present application.
Claims (10)
1. A method for extracting chenodeoxycholic acid from chicken bile paste comprises the steps of saponification, acid-adjusting extraction, methyl esterification, acetylation and reflux hydrolysis of the chicken bile paste, and is characterized by further comprising the step of silica gel filtration, wherein the silica gel filtration mode is that silica gel is paved in a filter funnel for filtration.
2. The method of claim 1, wherein the silica gel has a thickness of 2 to 5cm or 8 to 12 cm.
3. A method for extracting chenodeoxycholic acid from chicken bile paste comprises the following steps:
(1) saponification: adding sodium hydroxide and water into the chicken gall paste, heating, refluxing and saponifying to obtain a reaction solution;
(2) extraction and esterification: adding an ester solvent into the reaction liquid obtained in the step (1), adjusting acidity for extraction, concentrating the extract liquid until the reaction system becomes oily, adding methanol and concentrated sulfuric acid for reflux stirring, performing esterification, adding solid sodium bicarbonate to adjust the pH value after the esterification is finished, filtering to obtain a bile acid methyl ester methanol solution, and concentrating to obtain an oily substance;
(3) acetylation: adding an acetylation reagent into the oily substance obtained in the step (2) for reflux reaction, cooling to room temperature after the reaction is finished, filtering, and concentrating the filtrate to obtain an oily substance;
(4) silica gel filtration: adding an alkane solvent into the oily matter obtained in the step (3), heating to completely dissolve the oily matter obtained in the step (3), adding silica gel powder, cooling, spreading a layer of silica gel in a filter funnel for filtering, washing with the alkane solvent to obtain a filtrate, wherein a filter cake is triacetyl cholic acid methyl ester, and hydrolyzing the triacetyl cholic acid methyl ester recovered from the filter cake to obtain cholic acid;
(5) refluxing and hydrolyzing: concentrating and recovering the filtrate obtained in the step (4), adding a sodium hydroxide aqueous solution for reflux hydrolysis, cooling to adjust acidity, adding ethyl acetate for extraction, cooling to crystallize, and dropwise adding an alkane solvent to expel crystals to obtain chenodeoxycholic acid.
4. The method of claim 3, wherein the ester solvent added in step (2) is selected from one or more of methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, and isobutyl acetate.
5. The method of claim 3, wherein solid sodium bicarbonate is added in step (2) to adjust the pH to 7-8.
6. The process of claim 3, wherein in step (3) the acetylating reagent is selected from one of acetic anhydride, a combination of sodium acetate or a combination of acetyl chloride and triethylamine.
7. The method according to claim 3, wherein the mass of the silica gel powder added in the step (4) is 0.1-1 times of the mass of the chicken gall paste.
8. The method of claim 3, wherein the alkane solvent in step (4) and step (5) is one or more selected from hexane, heptane, dichloromethane, cyclohexane and petroleum ether.
9. Cholic acid and chenodeoxycholic acid prepared by the method of any one of claims 1 to 8.
10. Use of a process according to any one of claims 1 to 8 for the purification of chenodeoxycholic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911091094.7A CN110845564B (en) | 2019-11-14 | 2019-11-14 | Method for extracting chenodeoxycholic acid from chicken gall paste |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911091094.7A CN110845564B (en) | 2019-11-14 | 2019-11-14 | Method for extracting chenodeoxycholic acid from chicken gall paste |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110845564A true CN110845564A (en) | 2020-02-28 |
| CN110845564B CN110845564B (en) | 2022-09-16 |
Family
ID=69600981
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911091094.7A Active CN110845564B (en) | 2019-11-14 | 2019-11-14 | Method for extracting chenodeoxycholic acid from chicken gall paste |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110845564B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850846A (en) * | 2006-05-23 | 2006-10-25 | 辽宁百隆生物工程有限公司 | Production method for extracting chenodeoxycholic acid using chicken gall |
| CN101351470A (en) * | 2005-12-12 | 2009-01-21 | 株式会社大熊制药 | Purification process for chenodeoxycholic acid |
| CN106883281A (en) * | 2017-03-08 | 2017-06-23 | 眉山市新功生物科技有限公司 | The method that chenodeoxycholic acid is extracted from duck bile |
| CN107226832A (en) * | 2017-06-15 | 2017-10-03 | 马鞍山汇智生物技术有限公司 | A kind of new method that chenodeoxycholic acid is prepared by raw material of chicken courage |
| CN108218943A (en) * | 2018-03-05 | 2018-06-29 | 常德云港生物科技有限公司 | By the chenodeoxycholic acid in chicken courage and the method for cholic acid synthesis ursodesoxycholic acid |
-
2019
- 2019-11-14 CN CN201911091094.7A patent/CN110845564B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101351470A (en) * | 2005-12-12 | 2009-01-21 | 株式会社大熊制药 | Purification process for chenodeoxycholic acid |
| CN1850846A (en) * | 2006-05-23 | 2006-10-25 | 辽宁百隆生物工程有限公司 | Production method for extracting chenodeoxycholic acid using chicken gall |
| CN106883281A (en) * | 2017-03-08 | 2017-06-23 | 眉山市新功生物科技有限公司 | The method that chenodeoxycholic acid is extracted from duck bile |
| CN107226832A (en) * | 2017-06-15 | 2017-10-03 | 马鞍山汇智生物技术有限公司 | A kind of new method that chenodeoxycholic acid is prepared by raw material of chicken courage |
| CN108218943A (en) * | 2018-03-05 | 2018-06-29 | 常德云港生物科技有限公司 | By the chenodeoxycholic acid in chicken courage and the method for cholic acid synthesis ursodesoxycholic acid |
Non-Patent Citations (2)
| Title |
|---|
| 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》 * |
| 陈秋梦等: "鹅去胆酸提取分离", 《广东化工》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110845564B (en) | 2022-09-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110790805B (en) | Method for extracting chenodeoxycholic acid from pig bile paste | |
| CN102766185B (en) | Method for respectively recovering ursodesoxycholic acid and chenodeoxycholic acid from ursodesoxycholic acid waste mother liquor | |
| WO2012051774A1 (en) | Process for salting out and extracting organic acid from fermentation broth | |
| CN106674293B (en) | A method of sucralose-6-acetic ester waste mother liquor is handled using hydrolysis method | |
| CN1830996A (en) | Method for preparing chenodeoxycholic acid | |
| CN110256517B (en) | Method for producing high-purity chenodeoxycholic acid from pig bile or leftovers | |
| CN102453011A (en) | Preparation method of high-purity naringenin | |
| CN110845564B (en) | Method for extracting chenodeoxycholic acid from chicken gall paste | |
| CN110734467A (en) | method for extracting and purifying spinosad from fermentation liquor | |
| CN112062802A (en) | Chenodeoxycholic acid butyl acetate extracting solution and preparation method thereof, and chenodeoxycholic acid ammonium salt and chenodeoxycholic acid preparation method | |
| CN103087129B (en) | Method for extracting geniposide from gardenia yellow pigment waste liquor | |
| CN108484423B (en) | Method for separating and purifying L-alanine from L-alanine fermentation liquor | |
| JP2008043327A (en) | Method for producing succinic acid | |
| CN111995652B (en) | Method for separating chenodeoxycholic acid from duck bile | |
| CN109422794A (en) | A method of extracting chenodeoxycholic acid from duck bile | |
| CN111253458B (en) | Method for extracting stigmasterol from waste residues generated in natural ferulic acid production process | |
| CN108299538B (en) | Method for removing isoursodesoxycholic acid in duck bile | |
| CN106905145A (en) | A kind of preparation method of high-purity crocetin | |
| CN109053581B (en) | Preparation method of metronidazole benzoate | |
| CN111285915A (en) | Novel process for extracting and refining chenodeoxycholic acid from poultry gall bladder | |
| CN117801045A (en) | Method for separating and purifying three bile acids in pig bile | |
| CN106366038A (en) | Method of recycling dextromethorphan from crystallization mother liquid | |
| CN111635419B (en) | Method for treating cefdinir refined mother liquor | |
| CN116284171B (en) | Purification method of 4,1',6' -trichlorosucrose-6-acetate | |
| CN109400621B (en) | Preparation method of high-purity milbemycins |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |