CN112694486A - Solid-liquid separation method for tacrolimus fermentation liquor - Google Patents
Solid-liquid separation method for tacrolimus fermentation liquor Download PDFInfo
- Publication number
- CN112694486A CN112694486A CN202011563881.XA CN202011563881A CN112694486A CN 112694486 A CN112694486 A CN 112694486A CN 202011563881 A CN202011563881 A CN 202011563881A CN 112694486 A CN112694486 A CN 112694486A
- Authority
- CN
- China
- Prior art keywords
- tacrolimus
- resin
- solid
- liquid separation
- eluent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 title claims abstract description 37
- 229960001967 tacrolimus Drugs 0.000 title claims abstract description 36
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 title claims abstract description 36
- 238000000855 fermentation Methods 0.000 title claims abstract description 33
- 230000004151 fermentation Effects 0.000 title claims abstract description 33
- 238000000926 separation method Methods 0.000 title claims abstract description 22
- 239000007788 liquid Substances 0.000 title claims abstract description 17
- 239000011347 resin Substances 0.000 claims abstract description 44
- 229920005989 resin Polymers 0.000 claims abstract description 44
- 238000001179 sorption measurement Methods 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 10
- 239000000049 pigment Substances 0.000 claims abstract description 8
- 238000010828 elution Methods 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- 239000003480 eluent Substances 0.000 claims description 19
- 238000002386 leaching Methods 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 3
- 239000002904 solvent Substances 0.000 abstract description 6
- 230000007613 environmental effect Effects 0.000 abstract description 4
- 238000000605 extraction Methods 0.000 abstract description 4
- 238000000746 purification Methods 0.000 abstract description 4
- 239000003814 drug Substances 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000012535 impurity Substances 0.000 description 3
- 201000004681 Psoriasis Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 208000009137 Behcet syndrome Diseases 0.000 description 1
- 229930105110 Cyclosporin A Natural products 0.000 description 1
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 1
- 108010036949 Cyclosporine Proteins 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 241001647839 Streptomyces tsukubensis Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 208000004631 alopecia areata Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010322 bone marrow transplantation Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229960001265 ciclosporin Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 206010021198 ichthyosis Diseases 0.000 description 1
- 229940124452 immunizing agent Drugs 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 230000001861 immunosuppressant effect Effects 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 229920001470 polyketone Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 230000024053 secondary metabolic process Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
- C07D498/18—Bridged systems
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
Abstract
The invention belongs to the technical field of purification, and relates to a solid-liquid separation method of tacrolimus fermentation liquor, which comprises the following steps: (1) resin adsorption, (2) centrifugal separation, and (3) resin elution. The method firstly utilizes the density difference to centrifugally separate the mycelium from the macroporous adsorption resin, saves the solvent required by extraction, has high extract quality and less pigment, is more economical and reduces the environmental protection pressure.
Description
Technical Field
The invention relates to the technical field of purification, and particularly relates to a solid-liquid separation method for tacrolimus fermentation liquor.
Background
Tacrolimus (FK 506) is a novel twenty-three macrolide polyketone immunosuppressant extracted from fermentation broth of Streptomyces tsukubaensis No.9993 isolated from Japanese tsukubaensis soil in 1984 by Tacrolimus pharmaceutical industries. Clinical research shows that the immunosuppressive activity of the compound is 10-100 times of that of traditional immunizing agent cyclosporin A. The traditional Chinese medicine composition is widely applied to liver and bone marrow transplantation, prevention and treatment of rejection of drug-resistant kidney transplantation of other medicines, and treatment of various autoimmune diseases such as Behcet's disease, degenerative dermatitis, psoriasis, ichthyosis, alopecia areata, rheumatoid arthritis, psoriasis, multiple sclerosis, diabetes and the like. In the short time of coming into the market in China, the market share is rapidly increased, and the medicine becomes a first-line clinical medicine for rejection reaction after liver and kidney transplantation.
Tacrolimus is produced by fermentation, so that a lot of byproducts exist in the fermentation broth, and further purification is needed to separate the tacrolimus. The components in the fermentation liquor need to be subjected to solid-liquid separation, and the prior process steps are as follows:
step (1): and (3) supplementing macroporous adsorption resin accounting for 3% of the volume of the fermentation liquor into the fermentation liquor when the fermentation culture is carried out for 48 hours, adsorbing tacrolimus generated by secondary metabolism of mycelium by the macroporous adsorption resin in the culture process, and increasing the fermentation tank-placing unit by more than 20% compared with the unit without the resin.
Step (2): and after the fermentation is finished, performing solid-liquid separation on the fermentation liquor by using a plate-and-frame filter press, simultaneously collecting mycelium and adsorption resin, and after HPLC (high performance liquid chromatography) detection, finding that the content of tacrolimus in the mycelium is 0, wherein all the tacrolimus generated in the fermentation process is adsorbed by the resin.
And (3): the mycelia and the adsorbent resin were leached with 5-fold amount of methanol solution.
The process has the defects that mycelium and macroporous adsorption resin can be simultaneously collected when the tacrolimus fermentation liquor is filtered by a plate-and-frame filter press, and the mycelium and the macroporous adsorption resin cannot be effectively separated, so that the mycelium does not contain tacrolimus, the mycelium and the macroporous adsorption resin still need to be simultaneously leached during leaching, the solvent consumption in the leaching process is high, the impurities extracted by leaching are more, and a large amount of solvent post-treatment also generates great pressure on the environment.
Therefore, it is necessary to develop a new purification process to solve the above problems.
Disclosure of Invention
The invention mainly aims to provide a solid-liquid separation method for tacrolimus fermentation liquor, which can reduce the solvent required by extraction, improve the quality of an extract and reduce the environmental protection pressure.
The invention realizes the purpose through the following technical scheme: a solid-liquid separation method of tacrolimus fermentation liquor comprises the following steps:
(1) resin adsorption: adding macroporous adsorption resin accounting for 3% of the volume of the fermentation liquor into the fermentation liquor, and stirring until complete adsorption;
(2) centrifugal separation: performing solid-liquid separation on the fermentation liquor by using a high-speed tubular centrifuge, enabling the macroporous adsorption resin adsorbing tacrolimus to be positioned at the lower part of a centrifuge tube, and taking out the macroporous adsorption resin;
(3) resin elution: and eluting the pigment by using the macroporous adsorption resin with a first eluent, and then eluting the tacrolimus by using a second eluent to obtain a tacrolimus leaching liquor.
Specifically, the first eluent in the step (3) is a methanol solution with a 50% volume content.
Further, the amount of the first eluent used was 5 times the volume of the resin.
Specifically, the second eluent in the step (3) is a methanol solution with 75% volume content.
Further, the second eluent is used in an amount of 5 times the volume of the resin.
By adopting the technical scheme, the technical scheme of the invention has the beneficial effects that:
the method firstly utilizes the density difference to centrifugally separate the mycelium from the macroporous adsorption resin, saves the solvent required by extraction, has high extract quality and less pigment, is more economical and reduces the environmental protection pressure.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example (b):
the invention relates to a solid-liquid separation method of tacrolimus fermentation liquor, which comprises the following steps:
(1) resin adsorption: adding macroporous adsorption resin accounting for 3% of the volume of the fermentation liquor into the fermentation liquor, and stirring until complete adsorption;
(2) centrifugal separation: performing solid-liquid separation on the fermentation liquor by using a high-speed tubular centrifuge, enabling the macroporous adsorption resin adsorbing tacrolimus to be positioned at the lower part of a centrifuge tube, and taking out the macroporous adsorption resin;
(3) resin elution: and eluting the pigment by using the macroporous adsorption resin with a first eluent, and then eluting the tacrolimus by using a second eluent to obtain a tacrolimus leaching liquor.
The macroporous adsorption resin has good adsorption effect on tacrolimus, and can completely absorb the tacrolimus in mycelium, so that the active ingredients of the tacrolimus can be separated from the mycelium. The density difference exists between the macroporous absorption resin and the mycelium, so the macroporous absorption resin and the mycelium can be separated at low cost by using a centrifugal machine, the macroporous absorption resin with higher density is positioned at the lower part of the solution, then the solution with higher ratio is removed, and the macroporous absorption resin with lower treatment capacity and containing tacrolimus is left. The macroporous adsorption resin can also adsorb some useless pigment, so another eluent is needed to remove the pigment before the tacrolimus is eluted, and the tacrolimus leaching liquor with less impurities and high extraction rate can be obtained by eluting the remained part. Because only the macroporous adsorption resin part with less proportion is left after centrifugal separation, the amount of solvent needed by leaching is less, but the leaching liquor has high quality and less pigment, thereby being more economical and reducing the environmental protection pressure.
The first eluent in the step (3) is a methanol solution with the content of 50% by volume, and the dosage of the first eluent is 5 times of the volume of the resin. The second eluent in the step (3) is 75% methanol solution by volume, and the dosage of the second eluent is 5 times of the volume of the resin. The methanol has low cost and convenient recovery, and can meet the requirements of impurity removal and elution of tacrolimus under the condition of small using amount.
What has been described above are merely some embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made without departing from the inventive concept thereof, and these changes and modifications can be made without departing from the spirit and scope of the invention.
Claims (5)
1. A solid-liquid separation method of tacrolimus fermentation liquor comprises the following steps:
(1) resin adsorption: adding macroporous adsorption resin accounting for 3% of the volume of the fermentation liquor into the fermentation liquor, and stirring until complete adsorption;
(2) centrifugal separation: performing solid-liquid separation on the fermentation liquor by using a high-speed tubular centrifuge, enabling the macroporous adsorption resin adsorbing tacrolimus to be positioned at the lower part of a centrifuge tube, and taking out the macroporous adsorption resin;
(3) resin elution: and eluting the pigment by using the macroporous adsorption resin with a first eluent, and then eluting the tacrolimus by using a second eluent to obtain a tacrolimus leaching liquor.
2. The method for solid-liquid separation of tacrolimus fermentation broth according to claim 1, characterized in that: the first eluent in the step (3) is a methanol solution with 50% volume content.
3. The method for solid-liquid separation of tacrolimus fermentation broth according to claim 2, characterized in that: the amount of the first eluent was 5 times the volume of the resin.
4. The method for solid-liquid separation of tacrolimus fermentation broth according to claim 1, characterized in that: the second eluent in the step (3) is a methanol solution with 75% volume content.
5. The method for solid-liquid separation of tacrolimus fermentation broth according to claim 4, characterized in that: the second eluent was used in an amount of 5 times the volume of the resin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011563881.XA CN112694486A (en) | 2020-12-25 | 2020-12-25 | Solid-liquid separation method for tacrolimus fermentation liquor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011563881.XA CN112694486A (en) | 2020-12-25 | 2020-12-25 | Solid-liquid separation method for tacrolimus fermentation liquor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112694486A true CN112694486A (en) | 2021-04-23 |
Family
ID=75510737
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011563881.XA Pending CN112694486A (en) | 2020-12-25 | 2020-12-25 | Solid-liquid separation method for tacrolimus fermentation liquor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112694486A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113912624A (en) * | 2021-12-03 | 2022-01-11 | 北大方正集团有限公司 | Tacrolimus separation and purification method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101481715A (en) * | 2009-01-20 | 2009-07-15 | 南京工业大学 | Method for purifying tacrolimus through biological fermentation |
| CN102408435A (en) * | 2011-07-18 | 2012-04-11 | 南京工业大学 | Method for purifying ascomycin from streptomyces fermentation liquor |
| CN102936253A (en) * | 2012-11-12 | 2013-02-20 | 华北制药集团新药研究开发有限责任公司 | Preparation method of high-purity tacrolimus |
| CN106478664A (en) * | 2016-08-29 | 2017-03-08 | 广东蓝宝制药有限公司 | A kind of method of extraction purification tacrolimuss in fermentation liquid |
| CN108929335A (en) * | 2018-08-31 | 2018-12-04 | 福建省微生物研究所 | A kind of preparation method of tacrolimus coarse-grain |
-
2020
- 2020-12-25 CN CN202011563881.XA patent/CN112694486A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101481715A (en) * | 2009-01-20 | 2009-07-15 | 南京工业大学 | Method for purifying tacrolimus through biological fermentation |
| CN102408435A (en) * | 2011-07-18 | 2012-04-11 | 南京工业大学 | Method for purifying ascomycin from streptomyces fermentation liquor |
| CN102936253A (en) * | 2012-11-12 | 2013-02-20 | 华北制药集团新药研究开发有限责任公司 | Preparation method of high-purity tacrolimus |
| CN106478664A (en) * | 2016-08-29 | 2017-03-08 | 广东蓝宝制药有限公司 | A kind of method of extraction purification tacrolimuss in fermentation liquid |
| CN108929335A (en) * | 2018-08-31 | 2018-12-04 | 福建省微生物研究所 | A kind of preparation method of tacrolimus coarse-grain |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113912624A (en) * | 2021-12-03 | 2022-01-11 | 北大方正集团有限公司 | Tacrolimus separation and purification method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3896050B1 (en) | Method for preparing cannabidiol by means of high-speed countercurrent chromatography separation and purification | |
| EP0480040B1 (en) | Process for separating impurities from aqueous solution of crude ethanol | |
| CN102443012B (en) | A kind of method of purifying rapamycin from fermented liquid | |
| CN102936253A (en) | Preparation method of high-purity tacrolimus | |
| CN105732738B (en) | A kind of method of purification of tobramycin | |
| CN112694486A (en) | Solid-liquid separation method for tacrolimus fermentation liquor | |
| EP1697383B1 (en) | Process for the purification of tacrolimus | |
| CN110437059B (en) | Method for extracting pachymic acid A and pachymic acid B from Poria peel | |
| CN107417749B (en) | Resin filler separation method of coenzyme I | |
| CN113087723B (en) | Separation and purification method of sirolimus | |
| CN109406685B (en) | High performance liquid chromatography method for separating carfilzomib and isomers thereof | |
| CN1283636C (en) | Separation purification method of catechin monomer | |
| CN110590883B (en) | Method for extracting and separating spinosad from saccharopolyspora spinosa fermentation liquor by adopting resin | |
| CN113549061A (en) | Anisodine separated and purified from alkaloid extract and preparation method thereof | |
| CN101045718A (en) | Method for separating and purifying 10-deacetyl Baccatins III | |
| CN105420293A (en) | Method for separating and purifying resveratrol from traditional Chinese medicine polygonum cuspidatum extraction solution | |
| CN111909176B (en) | Method for recovering ascomycin and tacrolimus 8-propyl analogue from tacrolimus separation waste liquid | |
| CN101723927A (en) | Method for batch production, separation and purification of catechin monomers EGCG | |
| CN111547718A (en) | Composite activated carbon and application thereof in purifying tacrolimus | |
| CN108250217A (en) | A kind of method for preparing immunosuppressor sirolimus | |
| CN109232674B (en) | Method for extracting chrysin-8-C-beta-D-glucoside from Nanshan tea | |
| CN114516884B (en) | Purification method of high-purity tacrolimus | |
| CN114315892B (en) | Extraction method of soybean lecithin | |
| CN101712686B (en) | Method for separating and purifying tacrolimus in fermentation liquor | |
| CN109929000A (en) | A kind of feldamycin purification of products method |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210423 |
|
| RJ01 | Rejection of invention patent application after publication |