CN107619421B - The continuous chromatography isolation and purification method of lincomycin - Google Patents
The continuous chromatography isolation and purification method of lincomycin Download PDFInfo
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- CN107619421B CN107619421B CN201710852564.1A CN201710852564A CN107619421B CN 107619421 B CN107619421 B CN 107619421B CN 201710852564 A CN201710852564 A CN 201710852564A CN 107619421 B CN107619421 B CN 107619421B
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- OJMMVQQUTAEWLP-KIDUDLJLSA-N lincomycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@@H](C)O)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 OJMMVQQUTAEWLP-KIDUDLJLSA-N 0.000 title claims abstract description 57
- 229960005287 lincomycin Drugs 0.000 title claims abstract description 56
- OJMMVQQUTAEWLP-UHFFFAOYSA-N Lincomycin Natural products CN1CC(CCC)CC1C(=O)NC(C(C)O)C1C(O)C(O)C(O)C(SC)O1 OJMMVQQUTAEWLP-UHFFFAOYSA-N 0.000 title claims abstract description 47
- 238000004587 chromatography analysis Methods 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000000746 purification Methods 0.000 title claims abstract description 17
- 238000002955 isolation Methods 0.000 title claims abstract description 6
- 239000000047 product Substances 0.000 claims abstract description 68
- 239000003480 eluent Substances 0.000 claims abstract description 18
- 238000004519 manufacturing process Methods 0.000 claims abstract description 14
- 239000000706 filtrate Substances 0.000 claims abstract description 12
- POUMFISTNHIPTI-BOMBIWCESA-N hydron;(2s,4r)-n-[(1r,2r)-2-hydroxy-1-[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-methylsulfanyloxan-2-yl]propyl]-1-methyl-4-propylpyrrolidine-2-carboxamide;chloride Chemical compound Cl.CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@@H](C)O)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 POUMFISTNHIPTI-BOMBIWCESA-N 0.000 claims abstract description 6
- 229960001595 lincomycin hydrochloride Drugs 0.000 claims abstract description 6
- 238000000926 separation method Methods 0.000 claims abstract description 6
- 239000011347 resin Substances 0.000 claims description 84
- 229920005989 resin Polymers 0.000 claims description 84
- 239000007788 liquid Substances 0.000 claims description 55
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 54
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 40
- 239000000463 material Substances 0.000 claims description 34
- 238000001179 sorption measurement Methods 0.000 claims description 27
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 claims description 26
- 238000004140 cleaning Methods 0.000 claims description 18
- 238000010521 absorption reaction Methods 0.000 claims description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 12
- 239000002994 raw material Substances 0.000 claims description 11
- 230000000694 effects Effects 0.000 claims description 9
- 239000012535 impurity Substances 0.000 claims description 9
- 239000012141 concentrate Substances 0.000 claims description 8
- 238000001514 detection method Methods 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 8
- 239000002250 absorbent Substances 0.000 claims description 6
- 230000002745 absorbent Effects 0.000 claims description 6
- 238000013461 design Methods 0.000 claims description 6
- 238000011010 flushing procedure Methods 0.000 claims description 6
- 230000004907 flux Effects 0.000 claims description 6
- 238000004088 simulation Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- DZSDDKNXMARQMJ-AVXYAQEDSA-N (2S,4R)-4-ethyl-N-[(1R,2R)-2-hydroxy-1-[(2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-methylsulfanyloxan-2-yl]propyl]-1-methylpyrrolidine-2-carboxamide Chemical group CN1C[C@H](CC)C[C@H]1C(=O)N[C@H]([C@@H](C)O)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 DZSDDKNXMARQMJ-AVXYAQEDSA-N 0.000 claims description 3
- CWPMAVJRTHPUNS-UHFFFAOYSA-N Lincomycin B Natural products CCC1CC(NC(=O)C(C(C)O)C2OC(SC)C(O)C(O)C2O)N(C)C1 CWPMAVJRTHPUNS-UHFFFAOYSA-N 0.000 claims description 3
- DZSDDKNXMARQMJ-UHFFFAOYSA-N N-Desmethyllincomycin Natural products CN1CC(CC)CC1C(=O)NC(C(C)O)C1C(O)C(O)C(O)C(SC)O1 DZSDDKNXMARQMJ-UHFFFAOYSA-N 0.000 claims description 3
- GNPWCZOXLLXPNA-RETOTEICSA-N CCC[C@@H]1C[C@H](N(C)C1)C(=O)N[C@H]([C@@H](C)O)[C@H]1O[C@H](SCC)[C@H](O)[C@@H](O)[C@H]1O Chemical compound CCC[C@@H]1C[C@H](N(C)C1)C(=O)N[C@H]([C@@H](C)O)[C@H]1O[C@H](SCC)[C@H](O)[C@@H](O)[C@H]1O GNPWCZOXLLXPNA-RETOTEICSA-N 0.000 claims 1
- FHBOKMJKJSSKHN-UHFFFAOYSA-N Lincomycin C Natural products CCCC1CC(NC(=O)C(C(C)O)C2OC(SCC)C(O)C(O)C2O)N(C)C1 FHBOKMJKJSSKHN-UHFFFAOYSA-N 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 6
- 238000002360 preparation method Methods 0.000 abstract description 3
- 238000010828 elution Methods 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract 1
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- 238000011069 regeneration method Methods 0.000 abstract 1
- 150000003839 salts Chemical class 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 238000002425 crystallisation Methods 0.000 description 12
- 230000008025 crystallization Effects 0.000 description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
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- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241001655322 Streptomycetales Species 0.000 description 1
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- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The present invention relates to a kind of continuous chromatography isolation and purification method of lincomycin, the present invention realizes efficiently separating for Lincomycin A component using continuous chromatography chromatography technique.The present invention is by the fermented filtrate of lincomycin obtained by biofermentation, into continuous chromatography system through adsorbing, washing miscellaneous, elution lincomycin, collect eluent and chromatography column regeneration, the eluent concentration of collection, then through the Lincomycin Hydrochloride that decolourizes to obtain at salt.Lincomycin yield using this law separation is high, and purity is high, at low cost, and environmental protection is suitble to industrialized production.The present invention is short with the production technology time, product yield is high, the content of Lincomycin Hydrochloride (giving money as a gift) reaches 99% or more in the final products of preparation, the content of lincomycin hydrochloride for solving prior art production is not high, and the disadvantage of production technology time length.
Description
Technical field
The invention belongs to pharmaceutical fields, are related to a kind of isolation and purification method of antibacterials lincomycin.
Background technology
Lincomycin (C18H34N2O6S it) is also known as cillimycin, lincomycinum, is a kind of effective inhibition to gram-positive bacteria
Agent, action principle are to act on the ribosomal 50S subunits of sensitive bacteria, prevent the extension of peptide chain, inhibit the albumen of bacterial cell
Matter synthesizes.In higher concentrations, also there is bactericidal effect to highly sensitive bacterium.Lincomycin slow, group with resistance development
It knits good penetrability, infection in respiratory system etc. is had a better effect, therefore lincomycin is widely used in clinical treatment.In mould
The drug-fast bacteria of other antibiotic such as element is gradually found, and in the case of causing these antibiotic usages restricted, lincomycin needs
The amount of asking is gradually increasing.
It is divided into six kinds of components (being shown in Table 1) of A, B, C, D, K, S because of the difference of substituent group R in lincomycin molecule, is sending out
There are different degrees of expression, wherein component A to account for 90% or more during ferment, B component accounts for 5~10%, and clinical efficacy is only A groups
The 20% of part, and be more toxic, the total amount of remaining 4 kinds of component is no more than 1%, and curative effect is similar to component A.The derivative of B component
Clinical application have prodigious side effect, be especially affected to intestines, Liver and kidney function.Therefore, Lincomycin B component is at it
It is occupied an important position in quality control.In directly as medicinal lincomycin product, it is desirable that B component content be less than 5%, and
When making raw material production derivative using lincomycin, it is generally desirable to B component contents to be less than 3%.U.S. FDA export-oriented is even more to require
B component content is less than 1%.
1 lincomycin each component substituent group of table
By research it was found that the production technology of lincomycin is more traditional at present, the hydrochloric acid woods produced can be mould
Cellulose content is not high, and impurity content is also more, and the production technology time is long, cannot be satisfied the market demand.
The production technology of existing lincomycin：Lincomycin generally use lincomycin streptomycete under certain condition into
Row three grade fermemtation, is filtered to remove mycelium after the acidified processing of zymotic fluid, butanol is added with NaOH tune pH to 11 in filtrate thereto
Extraction refinement is carried out, the back extraction of HCL solution is added, after reduced pressure, activated carbon adsorption, crystallization, drying obtain Lincomycin Hydrochloride original
Expect medicine.
For this purpose, the present invention is improved on purification process, by continuous chromatography column on fermented filtrate of lincomycin, utilize
Adsorption rate of the lincomycin difference component on resin is different, achievees the purpose that isolate and purify, obtains a kind of woods of high-purity
Can mycin prepare method.
Invention content
The purpose of the present invention is to provide a kind of continuous chromatography isolation and purification methods of lincomycin.
Purification process of the present invention, steps are as follows：
Step 1, continuous chromatography column on fermented filtrate of lincomycin, obtains eluent；
Step 2, eluent obtains lincomycin sterling by concentration, crystallizing and drying.
Wherein, upper continuous chromatography column, resin described in step 1 are selected from：Macroporous absorbent resin HZ-802, R-802, Lan Xin -30,
II resin of AB-8, YPR-, resin particle diameter is in 50~100 mesh, 95% or more the uniformity.
Wherein, eluent described in step 2 is as follows by concentration, dry process：Concentration, which uses, to be concentrated under reduced pressure, and vacuum degree is
0.02~0.15Mpa, operation temperature are 35~90 DEG C, and it is 25%~70% to be concentrated into lincomycin mass concentration, is concentrated
Liquid；Concentrate obtains solid with the method that acetone crystallizes.
Wherein, continuous chromatography is selected from：Disk conveying type continuous chromatography piece-rate system or Simulation moving bed formula continuous chromatography
Piece-rate system.
Wherein, upper continuous chromatography column described in step 1, by following paragraphs district's groups at：
A, feed zone (14-17#)：
Charging is divided into two groups of parallel feeds in the region.By flow control, raw material initially enters 14# mouthfuls and 15# mouthfuls,
17# mouthfuls and 16# mouthfuls of series connection, material liquid passes through resin column, and product is attracted on resin, detects 14# column outlets feed liquid and raw material
Liquid is consistent, then is determined as supersaturated absorption.17# mouthfuls are middle material with 16# mouthfuls of feed liquids out, carry out applying mechanically charging.
B, water wash zone (10-13#) after charging：
4 valve ports in the region connect in a series arrangement, after being located at adsorption zone, after resin column adsorption saturation, turn to
There is excessive material liquid in the areas Xi Liao in resin column, are eluted remaining feed liquid in pillar and impurity B, C with water.It elutes
Feed liquid enters middle material and carries out applying mechanically charging.
C, secondary feeds area (18-20#)
The valve port in the region is to be connected in series with.Since adsorption zone supersaturation is adsorbed, part material liquid, institute are also contained in the areas Xi Liao
It secondary feeds area is arranged, can ensure the complete absorption of product in this way, increase product yield.Simultaneously because impurity B can A with woods
Adsorption rate on resin is different, is adjusted by flow velocity, can be eluted out impurity B, but also can take part woods out of can
A。
D, product parsing area (7-9#)
In the area, using being connected in series with mode.Behind the areas Xi Liao, the impurity B, C in resin column are seldom, make
Product completion is parsed with n-butanol.The region soon rotates previous minute in pillar, needs to detect the outlets 7#, really
It protects the outlets 7# and does not contain the product A or A of product containing minute quantity, then representative products parse completely.Another detection production life cycle sample, confirms solution
The product design and purity analysed.
E, the areas Xi Chun (2-6#)
In the area, using being connected in series with mode.Since n-butanol density ratio water is small, so cleaned using anti-water inlet,
Cleaning performance can be relatively good in this way, reduces cleaning water consumption, which needs the wash water for detecting the outlets 2# 1 minute before rotation,
It detects whether containing butanol, to judge whether the areas Xi Chun clean up.
F, pressure area (1#) under resin
Since the areas Xi Chun are using anti-water inlet cleaning way, so resin can be got on by top, 1# columns will be under resin using water
It pushes back and falls, it is ensured that the using effect of next period charging.Use high flux flushing, since resin column cleans up, the area
It can be used repeatedly for domain water.
Preferably, purification process of the invention, includes the following steps：
Step 1, continuous chromatography column on fermented filtrate of lincomycin, obtains eluent；
Step 2, eluent obtains lincomycin sterling by concentration, crystallizing and drying；
Wherein, upper continuous chromatography column, resin described in step 1 are selected from：Macroporous absorbent resin HZ-802, R-802, Lan Xin -30,
II resin of AB-8, YPR-, resin particle diameter is in 50~100 mesh, 95% or more the uniformity.Wherein, it is concentrated described in step 2 dry
Dry, method is as follows：Using being concentrated under reduced pressure, vacuum degree is 0.02~0.15Mpa for concentration, and operation temperature is 35~90 DEG C, is concentrated into
Lincomycin mass concentration is 25%~70%, obtains concentrate；Concentrate obtains solid with the method that acetone crystallizes；
Using the separation of disk conveying type continuous chromatography or the separation of Simulation moving bed formula continuous chromatography, steps are as follows：
A, feed zone (14-17#)：
Charging is divided into two groups of parallel feeds in the region.By flow control, raw material initially enters 14# mouthfuls and 15# mouthfuls,
17# mouthfuls and 16# mouthfuls of series connection, material liquid passes through resin column, and product is attracted on resin, detects 14# column outlets feed liquid and raw material
Liquid is consistent, then is determined as supersaturated absorption.17# mouthfuls are middle material with 16# mouthfuls of feed liquids out, carry out applying mechanically charging.
B, water wash zone (10-13#) after charging：
4 valve ports in the region connect in a series arrangement, after being located at adsorption zone, after resin column adsorption saturation, turn to
There is excessive material liquid in the areas Xi Liao in resin column, are eluted remaining feed liquid in pillar and impurity B, C with water.It elutes
Feed liquid enters middle material and carries out applying mechanically charging.
C, secondary feeds area (18-20#)
The valve port in the region is to be connected in series with.Since adsorption zone supersaturation is adsorbed, part material liquid, institute are also contained in the areas Xi Liao
It secondary feeds area is arranged, can ensure the complete absorption of product in this way, increase product yield.Simultaneously because impurity B can with woods
Adsorption rate of the mycin A on resin is different, is adjusted by flow velocity, can be eluted out impurity B；
D, product parsing area (7-9#)
In the area, using being connected in series with mode.Behind the areas Xi Liao, the impurity B, C in resin column are seldom, make
Product completion is parsed with n-butanol.The region soon rotates previous minute in pillar, needs to detect the outlets 7#, really
It protects the outlets 7# and does not contain the product A or A of product containing minute quantity, then representative products parse completely.Another detection production life cycle sample, confirms solution
The product design and purity analysed.
E, the areas Xi Chun (2-6#)
In the area, using being connected in series with mode.Since n-butanol density ratio water is small, so cleaned using anti-water inlet,
Cleaning performance can be relatively good in this way, reduces cleaning water consumption, which needs the wash water for detecting the outlets 2# 1 minute before rotation,
It detects whether containing butanol, to judge whether the areas Xi Chun clean up.
F, pressure area (1#) under resin
Since the areas Xi Chun are using anti-water inlet cleaning way, so resin can be got on by top, 1# columns will be under resin using water
It pushes back and falls, it is ensured that the using effect of next period charging.Using high flux flushing, resin column is cleaned up, the region
It can be used repeatedly for water.
Wherein, the impurity B, C are respectively Lincomycin B component；Other poly-doped impurities (C containing lincomycin, D, K, S and
Other).
Wherein, fermented filtrate of lincomycin is provided by Jiangxi Guoyao Co., Ltd.
On fermented filtrate of lincomycin before continuous chromatography column, Lincomycin A constituent content：20-25g/L, impurity woods can be mould
Plain B component content：4-4.5%, pH：10-10.5 optically-active detects：4.0-4.3.
Can effectively remove or reduce related impurities with the purification process of the present invention, it is obtained the result is that：Impurity woods
Can mycin B component≤0.3%, with high performance liquid chromatography (HPLC) measure Lincomycin Hydrochloride (Lincomycin A) purity >=
99.5%, moisture≤5%.
The continuous chromatography of the present invention isolates and purifies for existing method, has the following advantages：
1) shorten purification time, 40min is shorten to by original 8h；
2) cost is reduced, operating process is slowed down；
3) purity for improving lincomycin, reduces the generation of impurity；
4) due to continuous operation, product composition and concentration after absorption or ion exchange keep stablizing.
5) it is relatively fixed bed system, resin demand can reduce 50-90%.
Description of the drawings
Fig. 1 continuous chromatographies isolate and purify the flow chart of lincomycin
Fig. 2 embodiments 1：9-11 days product purities
Fig. 3 embodiments 2：22-26 days product purities
Fig. 4 embodiments 2：26-28 days product purities
Specific implementation mode
Embodiment 1
Step 1, continuous chromatography column on fermented filtrate of lincomycin, obtains eluent；
Step 2, eluent obtains lincomycin sterling by concentration, crystallizing and drying.
Wherein, upper continuous chromatography column, resin described in step 1 are selected from：Macroporous absorbent resin HZ-802, resin particle diameter exist
50~100 mesh, 95% or more the uniformity.
Wherein, eluent described in step 2 is as follows by concentration, dry process：Concentration, which uses, to be concentrated under reduced pressure, and vacuum degree is
0.02~0.15Mpa, operation temperature are 35~90 DEG C, and it is 25%~70% to be concentrated into lincomycin mass concentration, is concentrated
Liquid；Concentrate obtains solid with the method that acetone crystallizes.
Wherein, continuous chromatography is selected from：Disk conveying type continuous chromatography piece-rate system or Simulation moving bed formula continuous chromatography
Piece-rate system.
Wherein, upper continuous chromatography column described in step 1, by following paragraphs district's groups at：
A, feed zone (14-17#)：
Charging is divided into two groups of parallel feeds in the region.By flow control, raw material initially enters 14# mouthfuls and 15# mouthfuls,
17# mouthfuls and 16# mouthfuls of series connection, material liquid passes through resin column, and product is attracted on resin, detects 14# column outlets feed liquid and raw material
Liquid is consistent, then is determined as supersaturated absorption.17# mouthfuls are middle material with 16# mouthfuls of feed liquids out, carry out applying mechanically charging.
B, water wash zone (10-13#) after charging：
4 valve ports in the region connect in a series arrangement, after being located at adsorption zone, after resin column adsorption saturation, turn to
There is excessive material liquid in the areas Xi Liao in resin column, are eluted remaining feed liquid in pillar and impurity B, C with water.It elutes
Feed liquid enters middle material and carries out applying mechanically charging.
C, secondary feeds area (18-20#)
The valve port in the region is to be connected in series with.Since adsorption zone supersaturation is adsorbed, part material liquid, institute are also contained in the areas Xi Liao
It secondary feeds area is arranged, can ensure the complete absorption of product in this way, increase product yield.Simultaneously because impurity B can A with woods
Adsorption rate on resin is different, is adjusted by flow velocity, can be eluted out impurity B, but also can take part woods out of can
A。
D, product parsing area (7-9#)
In the area, using being connected in series with mode.Behind the areas Xi Liao, the impurity B, C in resin column are seldom, make
Product completion is parsed with n-butanol.The region soon rotates previous minute in pillar, needs to detect the outlets 7#, really
It protects the outlets 7# and does not contain the product A or A of product containing minute quantity, then representative products parse completely.Another detection production life cycle sample, confirms solution
The product design and purity analysed.
E, the areas Xi Chun (2-6#)
In the area, using being connected in series with mode.Since n-butanol density ratio water is small, so cleaned using anti-water inlet,
Cleaning performance can be relatively good in this way, reduces cleaning water consumption, which needs the wash water for detecting the outlets 2# 1 minute before rotation,
It detects whether containing butanol, to judge whether the areas Xi Chun clean up.
F, pressure area (1#) is since the areas Xi Chun are using anti-water inlet cleaning way under resin, so resin can be got on by top, in 1#
Column, which is pushed resin using water, to be fallen after rise, it is ensured that the using effect of next period charging.Using high flux flushing, due to resin
Column cleans up, and it can be used repeatedly for the regional water.
It is described in detail with reference to Fig. 1, Fig. 2 and embodiment
Feeding liquid is Jiangxi traditional Chinese medicines at present into the crude product before fixed bed：
Daily switch on condition and testing result：
| Date | Feed liquid purity (%) | Primary crystallization (%) | Secondary crystallization (%) |
| No. 9 | 99.12 | 99.52 | 99.54 |
| No. 10-11 | 99.26 | 99.57 | 99.63 |
Device parameter is as follows：
It is analyzed by data above, product purity can be stablized 99% or more.By September 10 days-September product on the 11st
Sampling is crystallized, and show that the product purity after primary crystallization is 99.57%, the product after secondary crystallization is 99.63%.It can sentence
Surely it can reach the product purity of target using the technique.And carry out secondary experiment judgement product yield.
Embodiment 2
Steps are as follows：
Step 1, continuous chromatography column on fermented filtrate of lincomycin, obtains eluent；
Step 2, eluent obtains lincomycin sterling by concentration, crystallizing and drying.
Wherein, upper continuous chromatography column, resin described in step 1 are selected from：Macroporous absorbent resin HZ-802, resin particle diameter exist
50~100 mesh, 95% or more the uniformity.
Wherein, eluent described in step 2 is as follows by concentration, dry process：Concentration, which uses, to be concentrated under reduced pressure, and vacuum degree is
0.02~0.15Mpa, operation temperature are 35~90 DEG C, and it is 25%~70% to be concentrated into lincomycin mass concentration, is concentrated
Liquid；Concentrate obtains solid with the method that acetone crystallizes.
Wherein, continuous chromatography is selected from：Disk conveying type continuous chromatography piece-rate system or Simulation moving bed formula continuous chromatography
Piece-rate system.
Wherein, upper continuous chromatography column described in step 1, by following paragraphs district's groups at：
A, feed zone (14-17#)：
Charging is divided into two groups of parallel feeds in the region.By flow control, raw material initially enters 14# mouthfuls and 15# mouthfuls,
17# mouthfuls and 16# mouthfuls of series connection, material liquid passes through resin column, and product is attracted on resin, detects 14# column outlets feed liquid and raw material
Liquid is consistent, then is determined as supersaturated absorption.17# mouthfuls are middle material with 16# mouthfuls of feed liquids out, carry out applying mechanically charging.
B, water wash zone (10-13#) after charging：
4 valve ports in the region connect in a series arrangement, after being located at adsorption zone, after resin column adsorption saturation, turn to
There is excessive material liquid in the areas Xi Liao in resin column, are eluted remaining feed liquid in pillar and impurity B, C with water.It elutes
Feed liquid enters middle material and carries out applying mechanically charging.
C, secondary feeds area (18-20#)
The valve port in the region is to be connected in series with.Since adsorption zone supersaturation is adsorbed, part material liquid, institute are also contained in the areas Xi Liao
It secondary feeds area is arranged, can ensure the complete absorption of product in this way, increase product yield.Simultaneously because impurity B can A with woods
Adsorption rate on resin is different, is adjusted by flow velocity, can be eluted out impurity B, but also can take part woods out of can
A。
D, product parsing area (7-9#)
In the area, using being connected in series with mode.Behind the areas Xi Liao, the impurity B, C in resin column are seldom, make
Product completion is parsed with n-butanol.The region soon rotates previous minute in pillar, needs to detect the outlets 7#, really
It protects the outlets 7# and does not contain the product A or A of product containing minute quantity, then representative products parse completely.Another detection production life cycle sample, confirms solution
The product design and purity analysed.
E, the areas Xi Chun (2-6#)
In the area, using being connected in series with mode.Since n-butanol density ratio water is small, so cleaned using anti-water inlet,
Cleaning performance can be relatively good in this way, reduces cleaning water consumption, which needs the wash water for detecting the outlets 2# 1 minute before rotation,
It detects whether containing butanol, to judge whether the areas Xi Chun clean up.
F, pressure area (1#) under resin
Since the areas Xi Chun are using anti-water inlet cleaning way, so resin can be got on by top, 1# columns will be under resin using water
It pushes back and falls, it is ensured that the using effect of next period charging.Use high flux flushing, since resin column cleans up, the area
It can be used repeatedly for domain water.
It is described in detail with reference to Fig. 1, Fig. 3, Fig. 4 and embodiment
Feeding liquid is Jiangxi traditional Chinese medicines at present into the crude product before fixed bed：
Daily switch on condition and testing result：
Secondary experiment September proceeds by the 22nd, purity data variation when following table is adjustment operation.
As can be seen from the above table：Previous experiments work-in-process purity is not high, and waste port has many products to come out, may
It is to cause feed liquid to be accumulated since secondary feeds region is long, the later stage reduces secondary feeds region pillar quantity, and product purity is gradual
Rise.After data stabilization, No. 26 start continuous feeds, judgment experiment stability and yield.Specific data are as follows：
Every batch of charging is fixed, and conservation of matter and calculated yield, specific data such as following table are done：
Yield is the sum of alcohol phase and water phase percentage.
Specific device parameter is as follows：
Its secondary feed zone pillar number is 2.
Sampling crystallization, experimental result are as follows：
| Date | Feed liquid purity (%) | Primary crystallization (%) | Secondary crystallization (%) |
| No. 27 | 99.53 | 99.68 | 99.74 |
| No. 28 first | 99.58 | 99.74 | 99.79 |
| No. 28 second batchs | 99.6 | 99.71 | 99.77 |
On the basis of 1 technique of embodiment, by adjusting different zones pillar quantity distribution ratio, seek to improve product purity
Optimal allocation ratio, and carry out product yield judgement.
The comparison of the purification process and existing purification process of embodiment 3, the present invention
In order to investigate the best preparation method of lincomycin, the preparation method of the present invention is compared with prior art
Experiment, wherein：Scheme 1：Be filtered to remove mycelium after the acidified processing of zymotic fluid, filtrate with NaOH tune pH to 11, thereto plus
Enter butanol and carry out extraction refinement, the back extraction of HCL solution is added, after reduced pressure, activated carbon adsorption, crystallization, drying, obtaining hydrochloric acid woods can
Mycin bulk pharmaceutical chemicals.Scheme 2：The purification process (embodiment 2) of the present invention.The result shows that the woods of purification process purifying of the invention
Can mycin purity index be substantially better than existing purification process.
Embodiment 4,
By purification process of the present invention, by feed liquid by load different model, particle diameter, the uniformity resin company
Continuous chromatographic column elution, later crystallization condition is identical, and gained lincomycin finished product purity data is as follows：
| Resin model | Particle diameter | The uniformity (%) | Feed liquid purity (%) | Primary crystallization (%) | Secondary crystallization (%) |
| HZ-802 | 50~100 | ≥95 | 99.53 | 99.68 | 99.74 |
| HZ-802 | ＞ 100 | ≥95 | 99.53 | 85.94 | 98.12 |
| HZ-802 | ＞ 100 | ＜ 95 | 99.53 | 84.72 | 96.65 |
| AB-8 | 50~100 | ≥95 | 99.53 | 99.52 | 99.69 |
| AB-8 | ＞ 100 | ≥95 | 99.53 | 86.15 | 97.95 |
| AB-8 | ＞ 100 | ＜ 95 | 99.53 | 83.18 | 96.35 |
| Lan Xin -30 | 50~100 | ≥95 | 99.53 | 99.45 | 99.66 |
| Lan Xin -30 | 50~100 | ＜ 95 | 99.53 | 86.35 | 98.37 |
| Lan Xin -30 | ＞ 100 | ＜ 95 | 99.53 | 84.17 | 96.28 |
| D-101 | 50~100 | ≥95 | 99.53 | 61.84 | 65.58 |
| D-101 | 50~100 | ＜ 95 | 99.53 | 59.75 | 62.38 |
| D-101 | ＞ 100 | ＜ 95 | 99.53 | 55.13 | 58.95 |
| BS-75 | 50~100 | ≥95 | 99.53 | 46.38 | 66.29 |
| BS-75 | 50~100 | ＜ 95 | 99.53 | 44.68 | 60.37 |
| BS-75 | ＞ 100 | ＜ 95 | 99.53 | 34.98 | 54.76 |
Claims (3)
1. a kind of continuous chromatography isolation and purification method of lincomycin, which is characterized in that steps are as follows：
Step 1, continuous chromatography column on fermented filtrate of lincomycin, obtains eluent；
Step 2, eluent obtains lincomycin sterling by concentration, crystallizing and drying；
Wherein, upper continuous chromatography column, resin described in step 1 are selected from：Macroporous absorbent resin HZ-802, R-802, Lan Xin -30, AB-
8, II resins of YPR-, resin particle diameter is in 50~100 mesh, 95% or more the uniformity, wherein is concentrated and dried described in step 2, side
Method is as follows：Using being concentrated under reduced pressure, vacuum degree is 0.02~0.15Mpa for concentration, and operation temperature is 35~90 DEG C, and being concentrated into woods can be mould
Quality amount a concentration of 25%~70%, obtains concentrate；Concentrate obtains solid with the method that acetone crystallizes；
Using the separation of disk conveying type continuous chromatography or the separation of Simulation moving bed formula continuous chromatography, steps are as follows：
A, feed zone (14-17#)：
Charging is divided into two groups of parallel feeds in the region, and by flow control, raw material initially enters 14# mouthfuls and 15# mouthfuls, is connecting
17# mouthfuls and 16# mouthfuls, material liquid passes through resin column, and product is attracted on resin, detection 14# column outlets feed liquid and material liquid one
It causing, is then determined as supersaturated absorption, 17# mouthfuls are middle material with 16# mouthfuls of feed liquids out, carry out applying mechanically charging,
B, water wash zone (10-13#) after charging：
4 valve ports in the region connect in a series arrangement, after being located at adsorption zone, after resin column adsorption saturation, turn to and wash material
There is excessive material liquid in area in resin column, is eluted remaining feed liquid in pillar and impurity B, C with water, the feed liquid eluted
It carries out applying mechanically charging into middle material,
C, secondary feeds area (18-20#)
The valve port in the region is to be connected in series with, and since adsorption zone supersaturation is adsorbed, part material liquid is also contained in the areas Xi Liao, so setting
Secondary feeds area is set, can ensure the complete absorption of product in this way, increases product yield, simultaneously because impurity B and Lincomycin A
Adsorption rate on resin is different, is adjusted by flow velocity, can be eluted out impurity B；
D, product parsing area (7-9#)
In the area, using mode is connected in series with, behind the areas Xi Liao, the impurity B, C in resin column are seldom, using just
Butanol parses product completion, which soon rotates previous minute in pillar, needs to detect the outlets 7#, it is ensured that 7#
Outlet does not contain Lincomycin A or Lincomycin A containing minute quantity, then representative products parse completely, another to detect production life cycle sample, really
Recognize the product design and purity parsed,
E, the areas Xi Chun (2-6#)
In the area, using mode is connected in series with, since n-butanol density ratio water is small, so being cleaned using anti-water inlet, in this way
Cleaning performance can be relatively good, reduces cleaning water consumption, which needs the wash water for detecting the outlets 2#, detection 1 minute before rotation
Whether butanol is contained, to judge whether the areas Xi Chun clean up,
F, pressure area (1#) under resin
Since the areas Xi Chun will be pushed back in 1# columns so resin can be got on by top using anti-water inlet cleaning way using water under resin
It falls, it is ensured that the using effect of next period charging is cleaned up resin column using high flux flushing, which can
With Reusability,
Wherein, impurity B is Lincomycin B,
Wherein, impurity C is the poly-doped impurity containing lincomycin C, D, K, S and other impurities.
2. the method according to claim 1, which is characterized in that Lincomycin Hydrochloride purity >=99.5%, moisture≤5%.
3. the method according to claim 1, which is characterized in that steps are as follows：
Step 1, continuous chromatography column on fermented filtrate of lincomycin, obtains eluent；
Step 2, eluent obtains lincomycin sterling by concentration, crystallizing and drying；
Wherein, upper continuous chromatography column, resin described in step 1 are selected from：Macroporous absorbent resin HZ-802, resin particle diameter 50~
100 mesh, 95% or more the uniformity；
Wherein, eluent described in step 2 is as follows by concentration, dry process：Concentration is using reduced pressure, vacuum degree 0.02
~0.15Mpa, operation temperature are 35~90 DEG C, and it is 25%~70% to be concentrated into lincomycin mass concentration, obtains concentrate；It is dense
Contracting liquid obtains solid with the method that acetone crystallizes；
Wherein, continuous chromatography is selected from：Disk conveying type continuous chromatography piece-rate system or the separation of Simulation moving bed formula continuous chromatography
System；
Wherein, upper continuous chromatography column described in step 1, by following paragraphs district's groups at：
A, feed zone (14-17#)：
Charging is divided into two groups of parallel feeds in the region, and by flow control, raw material initially enters 14# mouthfuls and 15# mouthfuls, is connecting
17# mouthfuls and 16# mouthfuls, material liquid passes through resin column, and product is attracted on resin, detection 14# column outlets feed liquid and material liquid one
It causing, is then determined as supersaturated absorption, 17# mouthfuls are middle material with 16# mouthfuls of feed liquids out, carry out applying mechanically charging,
B, water wash zone (10-13#) after charging：
4 valve ports in the region connect in a series arrangement, after being located at adsorption zone, after resin column adsorption saturation, turn to and wash material
There is excessive material liquid in area in resin column, is eluted remaining feed liquid in pillar and impurity B, C with water, the feed liquid eluted
It carries out applying mechanically charging into middle material,
C, secondary feeds area (18-20#)
The valve port in the region is to be connected in series with, and since adsorption zone supersaturation is adsorbed, part material liquid is also contained in the areas Xi Liao, so setting
Set secondary feeds area, can ensure the complete absorption of product in this way, increase product yield, simultaneously because impurity B and woods can A setting
Adsorption rate on fat is different, is adjusted by flow velocity, and impurity B is eluted out, but can also take out of part woods can A,
D, product parsing area (7-9#)
In the area, using mode is connected in series with, behind the areas Xi Liao, the impurity B, C in resin column are seldom, using just
Butanol parses product completion, which soon rotates previous minute in pillar, needs to detect the outlets 7#, it is ensured that 7#
Outlet does not contain Lincomycin A or Lincomycin A containing minute quantity, then representative products parse completely, another to detect production life cycle sample, really
Recognize the product design and purity parsed,
E, the areas Xi Chun (2-6#)
In the area, using mode is connected in series with, since n-butanol density ratio water is small, so being cleaned using anti-water inlet, in this way
Cleaning performance can be relatively good, reduces cleaning water consumption, which needs the wash water for detecting the outlets 2#, detection 1 minute before rotation
Whether butanol is contained, to judge whether the areas Xi Chun clean up,
F, pressure area (1#) under resin
Since the areas Xi Chun will be pushed back in 1# columns so resin can be got on by top using anti-water inlet cleaning way using water under resin
It falls, it is ensured that the using effect of next period charging uses high flux flushing, since resin column cleans up, the regional water
It can be used repeatedly.
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