Method for purifying erythromycin thiocyanate
By filtration through activated carbon and celite combined with acetone extraction and salting crystallization, the residual problem in the erythromycin thiocyanate crystallization mother liquor was solved, and erythromycin thiocyanate purification was achieved with high purity and high yield, reducing resource waste and environmental pressure.
Patent Information
- Application Number
- CN202410006181.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-01-03
- Publication Date
- 2025-07-11
AI Technical Summary
In the prior art, the residual erythromycin thiocyanate content in the crystallized mother liquor is low, resulting in a decrease in the loss of active ingredients and yield, and an increase in resource waste and environmental protection pressure.
The combined technology of activated carbon adsorption and celite filled column filtration combined with the primary crystallization of acetone extract salt and secondary crystallization of the crystallization mother liquor was used to remove impurities through activated carbon adsorption, and further purification was used by celite filtration, and purity and yield were improved by combining acetone extraction and salting crystallization.
It effectively improves the purity and yield of erythromycin thiocyanate, reduces resource waste, reduces environmental protection pressure, and improves production efficiency.
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Figure BDA0004647229690000011
Abstract
Description
Technical Field
[0001] The present invention relates to a method for purifying erythromycin thiocyanate, belonging to the technical field of antibiotic extraction. Background Art
[0002] Erythromycin thiocyanate (referred to as erythromycin red) is the thiocyanate of erythromycin, belonging to macrolide antibiotics. It can be used in the production of human and veterinary drugs. Clinically, it is mainly used to treat Gram-positive bacterial infections, and also has antibacterial effects on Gram-negative bacteria such as Haemophilus influenzae, Bordetella pertussis, Neisseria gonorrhoeae, Brucella and Neisseria meningitidis. In recent years, the market share of its synthetic derivatives clarithromycin, roxithromycin and azithromycin has increased rapidly in the domestic market. As the synthetic raw material of erythromycin thiocyanate, the demand for erythromycin thiocyanate also shows an upward trend. To meet the market demand, current research mainly focuses on improving the strains and fermentation levels of erythromycin thiocyanate, and there are also some studies on extraction. The compound information of erythromycin thiocyanate is as follows:
[0003] Chinese name: Erythromycin Thiocyanate
[0004] English name: Erythromycin Thiocyanate
[0005] CAS No.: 7704-67-8
[0006] Molecular formula: C38H68N2O13S
[0007] Molecular weight: 793.02
[0008] The chemical structural formula is as follows
[0009]
[0010] At present, the main extraction method of erythromycin thiocyanate is solvent extraction method, and the extraction solvent used is butyl acetate, that is, extracting with butyl acetate from the fermentation filtrate under alkaline conditions, and then through other treatments, and finally obtaining the finished product by crystallization. The main disadvantage of this process is that there is still erythromycin thiocyanate in the mother liquor after crystallization of erythromycin thiocyanate, which makes the content of the active ingredient reach the standard but the yield is lost. However, because the content of erythromycin thiocyanate remaining in the crystallization mother liquor is relatively low, it is difficult to extract and recover qualified erythromycin thiocyanate from it.
[0011] Therefore, recovering the residual erythromycin thiocyanate from the crystallization mother liquor, avoiding waste of resources and reducing environmental protection pressure have become urgent problems to be solved at present. Summary of the Invention
[0012] The object of the present invention is to provide a purification method for erythromycin thiocyanate. By using the combined pretreatment technology of activated carbon adsorption and diatomite packed column filtration for decolorization and impurity removal, and then combining the methods of salting out with acetone extract for primary crystallization and secondary crystallization of the crystallization mother liquor, the technical problems of high impurity content and waste of residual erythromycin thiocyanate in the primary crystallization mother liquor are solved, and the purity and yield of the product are effectively improved.
[0013] In order to achieve the above object, the technical solution adopted by the present invention is as follows:
[0014] A purification method for erythromycin thiocyanate, comprising the following steps:
[0015] (1) Alkalization and filtration: Add an alkalizing reagent to the erythromycin fermentation broth to adjust the pH value to 8.0 - 9.0, and obtain the erythromycin alkalized solution after solid-liquid separation;
[0016] (2) Decolorization and purification: Add activated carbon to the alkalized solution obtained in step (1) and stir for adsorption, and then filter in combination with a diatomite packed column to obtain a decolorized solution;
[0017] (3) Extraction: Add acetone to the decolorized solution obtained in step (2) for extraction, and separate the phases to obtain an acetone extract;
[0018] (4) Primary crystallization by salting out: Add a thiocyanate to the extract obtained in step (3) for salting out reaction, stand for crystal growth, and obtain wet erythromycin thiocyanate I and an acetone crystallization mother liquor after solid-liquid separation;
[0019] (5) Secondary crystallization by salting out: Concentrate and salt out the acetone crystallization mother liquor obtained in step (4) for crystallization, and obtain wet erythromycin thiocyanate II after solid-liquid separation;
[0020] (6) Drying: Mix and dry the wet erythromycin thiocyanate I obtained in step (4) and the wet erythromycin thiocyanate II obtained in step (5) to obtain the finished product of erythromycin thiocyanate.
[0021] Among them, the alkalizing reagent in step (1) is a sodium hydroxide or potassium hydroxide solution, and the concentration of the alkalizing reagent is 15% - 20%.
[0022] Among them, step (1) further includes filtering the alkalized solution through a ceramic membrane.
[0023] Among them, the dosage of activated carbon in step (2) is 15% - 20% of the volume of the filtrate.
[0024] Among them, the dosage of diatomite in step (2) is 10% - 15% of the volume of the filtrate.
[0025] Among them, the dosage of acetone in step (3) is 2 - 4 times the volume of the decolorized solution.
[0026] Among them, the thiocyanate described in step (4) is sodium thiocyanate or potassium thiocyanate, and the concentration of the thiocyanate is 8-15%.
[0027] Among them, the concentration in step (5) is nanofiltration concentration, and the titer of the nanofiltration concentrate is 20,000-30,000 μg / ml.
[0028] Among them, the salt in the salting-out in step (5) is solid sodium chloride or potassium chloride, and the dosage of the salt and the mass-volume ratio of the concentrate is 1:3-5.
[0029] Among them, the temperature of the crystallization in step (5) is 0-5°C.
[0030] The technical solution of the present invention has at least the following beneficial technical effects:
[0031] 1. The combined use of activated carbon adsorption and diatomite-packed column filtration can achieve a good decolorization effect and remove impurities. Combining the double crystallization method of acetone extraction liquid salting crystallization and crystallization mother liquor salting-out crystallization method for acetone mother liquor can effectively increase the content of erythromycin thiocyanate while improving the yield;
[0032] 2. By the method of recycling erythromycin thiocyanate from the erythromycin thiocyanate crystallization mother liquor, the residual erythromycin thiocyanate in the crystallization mother liquor can be effectively recycled, avoiding the waste of the residual erythromycin in the primary crystallization mother liquor, and at the same time avoiding the environmental pollution caused by the secondary crystallization mother liquor, reducing the treatment cost of production waste liquid, avoiding the waste of resources and reducing the environmental protection burden;
[0033] 3. The content of erythromycin thiocyanate A in the present invention reaches more than 90%, and the yield reaches more than 88%. Specific Embodiments
[0034] The following examples are used to illustrate the present invention, but are not used to limit the scope of the present invention. Unless otherwise specified, the percentage "%" involved in the examples refers to the mass percentage; but for the percentage of the solution, unless otherwise specified, it refers to the number of grams of solute contained in 100 ml of the solution.
[0035] In the following examples and comparative examples, the erythromycin fermentation broth was taken from the erythromycin fermentation workshop of Ningxia Taiyixin Biotechnology Co., Ltd. The fermentation was carried out by microbial fermentation culture of Streptomyces erythreus for 7 days. The pH of the fermentation broth was 6.0-7.0, the temperature was 32°C-36°C, and the fermentation unit was 9000±500 μg / ml.
[0036] In the following examples and comparative examples, the component content of erythromycin thiocyanate was detected by liquid phase detection method. The specific detection conditions were:
[0037] Chromatographic column: waters XBridge-C18, 4.6mm * 250mm, 5μm; Detection wavelength: 215nm; Injection volume: 40μl, Column temperature: 35°C; Flow rate: 1.0ml / min. The mobile phase is 0.025mol / l dipotassium hydrogen phosphate buffer solution: acetonitrile = 6:4 (volume ratio).
[0038] Example 1
[0039] Take 50L of erythromycin fermentation broth, add 15% NaOH solution to adjust the pH to 8.0, and obtain the filtrate by plate and frame filtration; add activated carbon accounting for 15% of the filtrate volume to the obtained filtrate, stir and adsorb, and then filter through a packed column filled with diatomaceous earth accounting for 15% of the filtrate volume to obtain the decolorized liquid. Add acetone twice the volume of the obtained decolorized liquid to stir and extract, let it stand for layer separation and separation to obtain the extract; add sodium thiocyanate to the obtained extract to stir and make erythromycin form a salt, let it stand for crystal cultivation, and obtain wet erythromycin thiocyanate and acetone mother liquor after ceramic membrane filtration; the obtained acetone mother liquor enters the nanofiltration membrane for filtration and concentration, and the obtained concentrated liquid has a titer of 24982μg / ml; add 1 / 3 of sodium chloride solution to the obtained acetone mother liquor, lower the temperature to 0°C and carry out crystal cultivation for 30min, and obtain wet erythromycin thiocyanate after ceramic membrane filtration; the obtained wet erythromycin thiocyanate is mixed evenly and dried to obtain the finished product of erythromycin thiocyanate. After detection, the content of erythromycin A is 91.31%, and the yield is 88.18%.
[0040] Example 2
[0041] Take 50L of erythromycin fermentation broth, add 17% KOH solution to adjust the pH to 8.5, and obtain the filtrate by plate and frame filtration; add activated carbon accounting for 16% of the filtrate volume to the obtained filtrate, stir and adsorb, and then filter through a packed column filled with diatomaceous earth accounting for 13% of the filtrate volume to obtain the decolorized liquid. Add acetone three times the volume of the obtained decolorized liquid to stir and extract, let it stand for layer separation and separation to obtain the extract; add sodium thiocyanate to the obtained extract to stir and make erythromycin form a salt, let it stand for crystal cultivation, and obtain wet erythromycin thiocyanate and acetone mother liquor after ceramic membrane filtration; the obtained acetone mother liquor enters the nanofiltration membrane for filtration and concentration, and the obtained concentrated liquid has a titer of 27381μg / ml; add 1 / 4 of potassium chloride solution to the obtained acetone mother liquor, lower the temperature to 3°C and carry out crystal cultivation for 30min, and obtain wet erythromycin thiocyanate after ceramic membrane filtration; the obtained wet erythromycin thiocyanate is mixed evenly and dried to obtain the finished product of erythromycin thiocyanate. After detection, the content of erythromycin A is 90.89%, and the yield is 88.74%.
[0042] Example 3
[0043] Take 50 L of erythromycin fermentation broth, add 20% NaOH solution to adjust the pH to 9.0, and obtain the filtrate through plate-and-frame filtration; add activated carbon accounting for 20% of the filtrate volume to the obtained filtrate, stir and adsorb, then filter through a packed column filled with diatomaceous earth accounting for 10% of the filtrate volume to obtain the decolorized solution. Add acetone twice the volume of the obtained decolorized solution to the decolorized solution, stir and extract, let it stand for layering and separation to obtain the extract; add sodium thiocyanate to the obtained extract and stir to make erythromycin form a salt, let it stand for crystal cultivation, and obtain wet erythromycin thiocyanate and acetone mother liquor after ceramic membrane filtration; the obtained acetone mother liquor enters the nanofiltration membrane for filtration and concentration to obtain a concentrated solution with a titer of 26374 μg / ml; add 1 / 5 of sodium chloride solution to the obtained acetone mother liquor, lower the temperature to 5°C and carry out crystal cultivation for 30 min, and obtain wet erythromycin thiocyanate after ceramic membrane filtration; mix the obtained wet erythromycin thiocyanate evenly and dry it to obtain the finished product of erythromycin thiocyanate. After detection, the content of erythromycin thiocyanate A is 91.06%, and the yield is 88.02%.
[0044] Comparative Example 1
[0045] Take 50 L of erythromycin fermentation broth, add 15% NaOH solution to adjust the pH to 8.0, and obtain the filtrate through plate-and-frame filtration; add activated carbon accounting for 15% of the filtrate volume to the obtained filtrate, stir and adsorb, then filter through a packed column filled with diatomaceous earth accounting for 13% of the filtrate volume to obtain the decolorized solution. Add acetone four times the volume of the obtained decolorized solution to the decolorized solution, stir and extract, let it stand for layering and separation to obtain the extract; add sodium thiocyanate to the obtained extract and stir to make erythromycin form a salt, let it stand for crystal cultivation, and obtain wet erythromycin thiocyanate after ceramic membrane filtration. Dry the obtained wet erythromycin thiocyanate to obtain the finished product of erythromycin thiocyanate. After detection, the content of erythromycin thiocyanate A is 90.89%, and the yield is 84.05%.
[0046] Comparative Example 2
[0047] Take 50 L of erythromycin fermentation broth, add 15% NaOH solution to adjust the pH to 8.0, and obtain the filtrate through plate-and-frame filtration; adsorb the obtained filtrate with a resin column, after elution with an eluent, the eluent is concentrated by nanofiltration and then sodium thiocyanate is added to form salt and crystallize, and obtain wet erythromycin thiocyanate and acetone mother liquor after ceramic membrane filtration; the obtained acetone mother liquor enters the nanofiltration membrane for filtration and concentration to obtain a concentrated solution with a titer of 24982 μg / ml; add 1 / 3 of sodium chloride solution to the obtained acetone mother liquor, lower the temperature to 0°C and carry out crystal cultivation for 30 min, and obtain wet erythromycin thiocyanate after ceramic membrane filtration; mix the obtained wet erythromycin thiocyanate evenly and dry it to obtain the finished product of erythromycin thiocyanate. After detection, the content of erythromycin thiocyanate A is 86.4%, and the yield is 87.68%
[0048] Comparative Example 3
[0049] Take 50 L of erythromycin fermentation broth, add 15% NaOH solution to adjust the pH to 8.0, and obtain the filtrate through plate-and-frame filtration; the obtained filtrate is adsorbed by a resin column, after elution with an eluent, the eluent is concentrated by nanofiltration and then sodium thiocyanate is added to form salt and crystallize, and the wet product of erythromycin thiocyanate is obtained through ceramic membrane filtration; the obtained wet product of erythromycin thiocyanate is dried to obtain the finished product of erythromycin thiocyanate. After detection, the content of erythromycin A thiocyanate is 85.45%, and the yield is 83.39%.
Claims
1. A method for purifying erythromycin thiocyanate, characterized in that It includes the following steps: (1) Alkalinization and filtration: Add an alkalinizing reagent to the erythromycin fermentation broth to adjust the pH value to 8.0 - 9.0, and obtain the erythromycin alkalinized solution after solid-liquid separation; (2) Decolorization and purification: Add activated carbon to the alkalinized solution obtained in step (1), stir and adsorb, and then filter through a diatomite-packed column in combination to obtain the decolorized solution; (3) Extraction: Add acetone to the decolorized solution obtained in step (2) for extraction, and separate the phases to obtain the acetone extraction solution; (4) Salt formation and primary crystallization: Add a thiocyanate to the extraction solution obtained in step (3) for salt formation reaction and crystallization, and obtain the wet product I of erythromycin thiocyanate and the acetone crystallization mother liquor after solid-liquid separation; (5) Salting-out and secondary crystallization: Concentrate and salt out the acetone crystallization mother liquor obtained in step (4) for crystallization, and obtain the wet product II of erythromycin thiocyanate after solid-liquid separation; (6) Drying: Mix and dry the wet product I of erythromycin thiocyanate obtained in step (4) and the wet product II of erythromycin thiocyanate obtained in step (5) to obtain the finished product of erythromycin thiocyanate.
2. The method according to claim 1, characterized in that The alkalinizing reagent described in step (1) is sodium hydroxide or potassium hydroxide solution, and the concentration of the alkalinizing reagent is 15% - 20%.
3. The method according to claim 1, wherein Step (1) further includes filtering the alkalinized solution through a ceramic membrane.
4. The method according to claim 1, wherein The dosage of the activated carbon described in step (2) is 15% - 20% of the volume of the filtrate.
5. The method according to claim 1, characterized in that The dosage of the diatomite described in step (2) is 10 - 15% of the volume of the filtrate.
6. The method according to claim 1, characterized in that The dosage of the acetone described in step (3) is 2 - 4 times the volume of the decolorized solution.
7. The method according to claim 1, characterized in that The thiocyanate described in step (4) is sodium thiocyanate or potassium thiocyanate, and the concentration of the thiocyanate is 8% - 15%.
8. The method according to claim 1, characterized in that The concentration in step (5) is nanofiltration concentration.
9. The method according to claim 8, wherein The titer of the nanofiltration concentrate is 20000 - 30000 μg / ml.
10. The method according to claim 1, characterized in that The salt in the salting-out described in step (5) is solid sodium chloride or potassium chloride, and the dosage of the salt is in a mass-volume ratio of 1:3 - 5 to the concentrate.