CN104327167A - Technology for extracting polymyxin B through precipitation method - Google Patents
Technology for extracting polymyxin B through precipitation method Download PDFInfo
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- CN104327167A CN104327167A CN201410510088.1A CN201410510088A CN104327167A CN 104327167 A CN104327167 A CN 104327167A CN 201410510088 A CN201410510088 A CN 201410510088A CN 104327167 A CN104327167 A CN 104327167A
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Abstract
The invention discloses a technology for extracting polymyxin B through a precipitation method. In the technology, by means of the precipitation method, a polymyxin B fermentation liquid is subjected to precipitation with oxalic acid, wherein a precipitated and separated solid is dissolved with addition of an alkaline substance and then is adsorbed by a weak acidic cation exchange resin. The weak acidic cation exchange resin is washed by purified water and is per-washed and desorbed with dilute sulfuric acid. A desorption solution is subjected to precipitation with an alkaline substance and acidic substances are dissolved. Solid-liquid separation is carried out with a liquid being subjected to decoloration and nano-filtration and then being dried to obtain the polymyxin B. The technology is short in required processes and is free of organic solvents. The polymyxin B is high in content and is suitable for industrialized production and application.
Description
Technical field
The invention belongs to medical art, be specifically related to a kind of technique adopting the precipitator method to extract PXB.
Background technology
Polymyxin is the alkaline ring type polypeptide with good anti-microbial activity being synthesized in metabolic process by bacillus polymyxa and secrete, PXB is the one of polymyxin, its vitriol conventional, for white crystalline powder, soluble in water, have draw moist, stablize in an acidic solution, its neutral solution is placed not affect for one week in room temperature and is tired, and basic solution is unstable, dissolves hardly in the organic solvents such as ether, ketone, ester, hydrocarbon.
PXB has stronger anti-microbial effect to most gram negative bacillus, more remarkable to the effect of Pseudomonas aeruginosa, also there is good effect to intestinal bacteria, Salmonellas, dysentery bacterium, hemophilus influenza, bordetella pertussis, gas bacillus and pneumobacillus etc.The bacterium of PXB to growth and breeding phase and stationary phase is all effective, past is mainly used in treating gram positive bacterial infection clinically, the particularly various infection that cause of Pseudomonas aeruginosa and intestinal bacteria, as respiratory system infection, peritonitis, bile duct infection, urinary tract infections, burn infection, cornea infect and septicemia etc.At present still can locally for the eye caused by sensitive organism, ear, skin, mucosal infections and burn charrin's disease.
PXB can be separated by fermentation liquor, extract, purifying, drying and obtaining.Patent GB742589, GB782926 etc. relate to and carry out absorption filtrate with weakly acidic cation-exchange resin, and desorbed solution continuation resin carries out purifying, and the method repeatedly uses resin, have the shortcomings such as production hour is long, absorption yield is lower.The patents such as GB9798887, CN103130875, CN103130876 are mentioned and are with an organic solvent carried out extracting the object reaching separation, and the method with an organic solvent, can cause certain injury, and environmental protection cost is higher to operator and environment.
Summary of the invention
The object of this invention is to provide that a kind of operation is short, content is high, the technique of environment amenable extraction PXB, solve the problem that in prior art, PXB extracting method operation is long, environmental pollution is serious, products obtained therefrom content is low.
In order to realize object of the present invention, inventor provide following technical scheme.
Adopt the precipitator method to extract a technique for PXB, comprise following operation steps:
A. the fermented liquid of PXB being added oxalic acid adjust ph is 1.0 ~ 2.0, carry out solid-liquid separation subsequently, 15 ~ 25 DEG C are cooled to by being separated the liquid obtained, adding alkaline matter adjust ph is 6.0 ~ 8.0, gained solution is transferred to weakly acidic cation-exchange resin post and adsorbs, be 80 ~ 100% by purified water washing resin post to the transparence of effluent liquid, then the sulfuric acid of 0.01 ~ 0.05mol/L is used to carry out prewashing to resin column, finally with the sulfuric acid of 0.1 ~ 0.5mol/L, resin column is resolved, collect desorbed solution;
B. adding alkaline matter adjust ph in the desorbed solution obtained to step a is 8.0 ~ 10.0, carry out solid-liquid separation subsequently, wash being separated the solid purified water obtained, purified water is added in the solid after washing, stir, then Plus acidic solution is until solid all dissolves, period control ph be 4.6 ~ 5.0, filter, gained filtrate is for subsequent use;
C. the filtrate obtained by step b carrying out is decoloured, nanofiltration, drying, namely obtain PXB.
Alkaline matter described in step a and step b is 3% ~ 7% aqueous sodium hydroxide solution.
Acidic substance described in step b are the sulfuric acid of 0.01 ~ 0.05mol/L.
15 ~ 20 DEG C are cooled to by being separated the liquid obtained in step a.
Be 98% ~ 100% by purified water washing resin post to the transparence of effluent liquid in step a.Can water-soluble impurity be removed, then pass through the sulfuric acid prewashing of 0.01 ~ 0.02mol/L, make the purity of desorbed solution reach more than 75%.
The sulfuric acid of 0.01 ~ 0.02mol/L is used to carry out prewashing to resin column in step a.
With the sulfuric acid of 0.1-0.2mol/L, resin column is resolved in step a.
Adding alkaline matter adjust ph in the desorbed solution obtained to step a in step b is 9.0 ~ 10.0.
Wash being separated the solid purified water obtained described in step b, the mass ratio of purified water and solid is (60 ~ 90): 1.
In the solid after washing, add purified water described in step b, the mass ratio of purified water and solid is (2 ~ 5): 1.
The technique of extraction PXB of the present invention, adopts the precipitator method, and the fermented liquid first to PXB adds oxalic acid adjust ph, object: 1. because the calcium ions and magnesium ions in oxalic acid and fermented liquid forms precipitation, can remove portion calcium ions and magnesium ions; 2. can by the removing of acidic protein precipitation after being adjusted to acidity.
After fermented liquid solid-liquid separation, it is 6.0 ~ 8.0 that filtrate adds alkaline matter adjust ph, and filtrate is adsorbed in neutral conditions.After absorption terminates, be 80 ~ 100% by purified water washing resin post to the transparence of effluent liquid, remove water miscible impurity.Then resolved by prewashing and obtain desorbed solution.
In desorbed solution, add alkaline matter adjust ph is 8.0 ~ 10.0, colistin sulfate B is precipitated, then precipitation purified water is washed, and mainly in order to remove the sodium ion wrapped up in precipitation, makes the feed liquid ash content after sulfuric acid dissolution keep qualified.
Add alkaline matter adjust ph precipitation and purified water washing in step b, the ash content of product can be made qualified, then by 0.01 ~ 0.05mol/L sulfuric acid dissolution, control ph is 4.6 ~ 5.0, vitriol and the PH of guarantee the finished product are qualified.
The fermented liquid that the fermented liquid of PXB of the present invention can adopt this area to commonly use bacterial classification to obtain after 48h fermentation.The technique of extraction PXB provided by the present invention, operation is short, simple to operate, whole process, without the need to an organic solvent, therefore can not cause damage to the healthy of operator, also can not to environment, the environmental protection pressure of manufacturing enterprise can be alleviated, the PXB content prepared is more than 88%, and economic benefits, is suitable for applying.
Embodiment
Below in conjunction with specific embodiment, content of the present invention is further described in detail.
Embodiment 1
A. the fermented liquid 200L of PXB, content 1.2mg/mL, adding oxalic acid adjust ph is 1.5, filters at normal temperatures, filtrate is cooled to 18 DEG C with ceramic membrane, adding 5% sodium hydroxide adjust ph is 6.9, gained solution is transferred to D152 resin column and adsorbs, and is 98%, then uses the sulfuric acid of 0.02mol/L to carry out prewashing to resin column by purified water washing resin post to the transparence of effluent liquid, finally with the sulfuric acid of 0.2mol/L, resin column is resolved, collect desorbed solution;
B. adding 5% sodium hydroxide adjust ph in the desorbed solution obtained to step a is 9.0, filter subsequently, obtain solid 410g (wet product), wash being separated the solid purified water 30L obtained, purified water 1000mL is added in the solid after washing, stir, make solid dispersed in purified water, then 0.03mol/L sulfuric acid is added until solid all dissolves, adition process control ph is 4.6-5.0, and when solid all dissolves, pH value is 4.65, stops adding sulfuric acid, filter, gained filtrate is for subsequent use;
C. the filtrate obtained by step b carrying out is decoloured, nanofiltration, drying, namely obtain PXB finished product 142g, content 90.3%, yield 67.5%.
Embodiment 2
A. the fermented liquid 250L of PXB, content 0.9mg/mL, adding oxalic acid adjust ph is 1.0, uses Plate Filtration at normal temperatures, is cooled to 17 DEG C by being separated the liquid obtained, adding 5% sodium hydroxide solution adjust ph is 6.0, gained solution is transferred to D152 resin column and adsorbs, and is 98.5%, then uses the sulfuric acid of 0.01mol/L to carry out prewashing to resin column by purified water washing resin post to the transparence of effluent liquid, finally with the sulfuric acid of 0.1mol/L, resin column is resolved, collect desorbed solution;
B. adding 5% sodium hydroxide adjust ph in the desorbed solution obtained to step a is 8.0, filtering-depositing, obtain solid 425g (wet product), washing being separated the solid purified water 35L obtained, in the solid after washing, adding purified water 1200mL, stir, make solid dispersed in purified water, then adding 0.05mol/L sulfuric acid control ph is 4.6-5.0, and when solid dissolves completely, pH value is 4.73, filter, gained filtrate is for subsequent use;
C. the filtrate obtained by step b carrying out is decoloured, nanofiltration, drying, namely obtain PXB finished product 130g, content 88.7%, yield 66.6%.
Embodiment 3
A. the fermented liquid 150L of PXB, content 1.6mg/mL, adding oxalic acid adjust ph is 2.0, filter with ceramic membrane at normal temperatures, solid-liquid separation, 15 DEG C are cooled to by being separated the liquid obtained, adding 6% sodium hydroxide solution adjust ph is 8.0, gained solution D152 resin column adsorbs, be 99% by purified water washing resin post to the transparence of effluent liquid, then use the sulfuric acid of 0.05mol/L to carry out prewashing to resin column, finally with the sulfuric acid of 0.5mol/L, resin column is resolved, collect desorbed solution;
B. adding 6% sodium hydroxide solution adjust ph in the desorbed solution obtained to step a is 10.0, filtering subsequently, obtain solid 450g (wet product), washing being separated the solid purified water 40L obtained, purified water 1500mL is added in the solid after washing, stir, make solid dispersed in purified water, then add the sulfuric acid of 0.04mol/L until solid all dissolves, adition process control ph is 4.6-5.0, when solid all dissolves, pH value is 4.72, and filter, gained filtrate is for subsequent use;
C. the filtrate obtained by step b carrying out is decoloured, nanofiltration, drying, namely obtain PXB finished product 150g, content 90.3%, yield 70.8%.
Claims (10)
1. adopt the precipitator method to extract a technique for PXB, it is characterized in that, comprise following operation steps:
A. the fermented liquid of PXB being added oxalic acid adjust ph is 1.0 ~ 2.0, carry out solid-liquid separation subsequently, 15 ~ 25 DEG C are cooled to by being separated the liquid obtained, adding alkaline matter adjust ph is 6.0 ~ 8.0, gained solution is transferred to weakly acidic cation-exchange resin post and adsorbs, be 80 ~ 100% by purified water washing resin post to the transparence of effluent liquid, then the sulfuric acid of 0.01 ~ 0.05mol/L is used to carry out prewashing to resin column, finally with the sulfuric acid of 0.1 ~ 0.5mol/L, resin column is resolved, collect desorbed solution;
B. adding alkaline matter adjust ph in the desorbed solution obtained to step a is 8.0 ~ 10.0, carry out solid-liquid separation subsequently, wash being separated the solid purified water obtained, purified water is added in the solid after washing, stir, then Plus acidic solution is until solid all dissolves, period control ph be 4.6 ~ 5.0, filter, gained filtrate is for subsequent use;
C. the filtrate obtained by step b carrying out is decoloured, nanofiltration, drying, namely obtain PXB.
2. a kind of technique adopting the precipitator method to extract PXB according to claim 1, it is characterized in that, alkaline matter described in step a and step b is 3% ~ 7% aqueous sodium hydroxide solution.
3. a kind of technique adopting the precipitator method to extract PXB according to claim 1, it is characterized in that, acidic substance described in step b are the sulfuric acid of 0.01 ~ 0.05mol/L.
4. a kind of technique adopting the precipitator method to extract PXB according to claim 1, is characterized in that, is cooled to 15 ~ 20 DEG C in step a by being separated the liquid obtained.
5. a kind of precipitator method that adopt according to claim 1 extract the technique of PXB, it is characterized in that, are 98% ~ 100% by purified water washing resin post to the transparence of effluent liquid in step a.
6. a kind of technique adopting the precipitator method to extract PXB according to claim 1, is characterized in that, use the sulfuric acid of 0.01 ~ 0.02mol/L to carry out prewashing to resin column in step a.
7. a kind of technique adopting the precipitator method to extract PXB according to claim 1, is characterized in that, resolve in step a with the sulfuric acid of 0.1-0.2mol/L to resin column.
8. a kind of precipitator method that adopt according to claim 1 extract the technique of PXB, it is characterized in that, adding alkaline matter adjust ph in the desorbed solution obtained to step a in step b is 9.0 ~ 10.0.
9. a kind of technique adopting the precipitator method to extract PXB according to claim 1, is characterized in that, wash being separated the solid purified water obtained described in step b, the mass ratio of purified water and solid is (60 ~ 90): 1.
10. a kind of technique adopting the precipitator method to extract PXB according to claim 1, it is characterized in that, in the solid after washing, add purified water described in step b, the mass ratio of purified water and solid is (2 ~ 5): 1.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018032506A1 (en) * | 2016-08-19 | 2018-02-22 | 湖北瑞昊安科医药科技发展有限公司 | Polymyxin b sulphate crystal and preparation method therefor |
| CN107759669A (en) * | 2016-08-19 | 2018-03-06 | 湖北瑞昊安科医药科技发展有限公司 | A kind of aerosporin crystallization and preparation method thereof |
| CN108977482A (en) * | 2018-08-21 | 2018-12-11 | 浙江日升昌药业有限公司 | A kind of preparation method of aerosporin |
Citations (8)
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|---|---|---|---|---|
| GB742589A (en) * | 1953-01-03 | 1955-12-30 | Distillers Co Yeast Ltd | Recovery of polymyxin |
| GB891534A (en) * | 1957-12-16 | 1962-03-14 | Hamao Umezawa | Kanamycin b and its isolation from a fermentation broth |
| GB991602A (en) * | 1961-03-10 | 1965-05-12 | Wellcome Found | Antibiotics and their purification |
| CN1800201A (en) * | 2005-12-06 | 2006-07-12 | 河北工业大学 | Polymyxin E separation preparation method |
| CN101974075A (en) * | 2010-10-12 | 2011-02-16 | 山东鲁抗医药股份有限公司 | Method for extracting polymyxin B and E from fermentation technique culture |
| CN102718842A (en) * | 2012-07-16 | 2012-10-10 | 河北圣雪大成制药有限责任公司 | Process for extracting colistin sulfate through precipitation method |
| CN103059105A (en) * | 2012-12-28 | 2013-04-24 | 三达膜科技（厦门）有限公司 | Method for extraction of colistin sulphate |
| CN103540633A (en) * | 2013-10-24 | 2014-01-29 | 河北圣雪大成制药有限责任公司 | Method for producing polymyxin B through fermentation process |
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2014
- 2014-09-28 CN CN201410510088.1A patent/CN104327167A/en active Pending
Patent Citations (8)
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| GB742589A (en) * | 1953-01-03 | 1955-12-30 | Distillers Co Yeast Ltd | Recovery of polymyxin |
| GB891534A (en) * | 1957-12-16 | 1962-03-14 | Hamao Umezawa | Kanamycin b and its isolation from a fermentation broth |
| GB991602A (en) * | 1961-03-10 | 1965-05-12 | Wellcome Found | Antibiotics and their purification |
| CN1800201A (en) * | 2005-12-06 | 2006-07-12 | 河北工业大学 | Polymyxin E separation preparation method |
| CN101974075A (en) * | 2010-10-12 | 2011-02-16 | 山东鲁抗医药股份有限公司 | Method for extracting polymyxin B and E from fermentation technique culture |
| CN102718842A (en) * | 2012-07-16 | 2012-10-10 | 河北圣雪大成制药有限责任公司 | Process for extracting colistin sulfate through precipitation method |
| CN103059105A (en) * | 2012-12-28 | 2013-04-24 | 三达膜科技（厦门）有限公司 | Method for extraction of colistin sulphate |
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Non-Patent Citations (1)
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| 张雪锋: "硫酸多黏菌素B的脱色工艺研究", 《畜牧与饲料科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018032506A1 (en) * | 2016-08-19 | 2018-02-22 | 湖北瑞昊安科医药科技发展有限公司 | Polymyxin b sulphate crystal and preparation method therefor |
| CN107759669A (en) * | 2016-08-19 | 2018-03-06 | 湖北瑞昊安科医药科技发展有限公司 | A kind of aerosporin crystallization and preparation method thereof |
| CN108977482A (en) * | 2018-08-21 | 2018-12-11 | 浙江日升昌药业有限公司 | A kind of preparation method of aerosporin |
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