Extraction method of adipyl-7-ADCA
By obtaining the adipyl-7-ADCA fermentation broth filtrate with mixed solvent and surfactant and adjusting the pH value, the turbulent state is achieved for extraction and separation, which solves the difficulty of adipicoyl-7-ADCA extraction in the prior art. The problem of this problem has achieved an efficient and safe extraction process and is suitable for large-scale production.
Patent Information
- Application Number
- CN202411333986.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-24
- Publication Date
- 2024-12-17
AI Technical Summary
There are few researches on the extraction and preparation process of adipicyl-7-ADCA in the prior art, and the lack of effective extraction methods makes it difficult to extract the important semi-synthetic intermediate of cephalosporin antibiotics.
The filtrate of adipicyl-7-ADCA fermentation broth is obtained by mixing and stirring with mixed solvents and surfactant, adjusting the pH value, and then extracting and separation after reaching the turbulent state, combining with environmentally friendly recycling treatment to improve the extraction efficiency and purity.
It achieves high yield and high purity extraction of adipicyl-7-ADCA, with a gentle process suitable for large-scale production and strong impurity removal ability.
Smart Images
Figure CN119143782A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of pharmaceutical biotechnology, and in particular to a method for extracting adipoyl-7-ADCA. Background Art
[0002] Cephalosporin is a general term for cephalosporins. Cephalosporins are a class of antibiotics obtained by semi-synthetically modifying the side chain of natural cephalosporin C obtained by culturing Cephalosporin C. Cephalosporin has a broad antibacterial spectrum, strong antibacterial activity, and low adverse reactions and toxic side effects. It has become the largest sales volume of antibiotic anti-infective drugs in the market.
[0003] Among cephalosporin products, 7-adipoylamino deacetoxy cephalosporanic acid (abbreviated as adipoyl-7-ADCA) is a relatively important API and an important semi-synthetic intermediate of cephalosporin antibiotics. It is mainly used in the synthesis of drugs with large market usage, such as cephalexin, cephradine and cefadroxil. Therefore, the extraction and preparation of adipoyl-7-ADCA is of great significance to the synthesis of cephalosporin drugs.
[0004] At present, researchers at home and abroad have rarely studied the extraction and preparation process of adipoyl-7-ADCA, and there are few related articles. Therefore, how to effectively extract adipoyl-7-ADCA is an urgent problem to be solved. It is particularly important to develop a simple, safe and reliable method for extracting adipoyl-7-ADCA. Summary of the invention
[0005] The embodiment of the present invention provides a method for extracting adipoyl-7-ADCA to solve the above technical problems in the prior art.
[0006] In order to achieve the above technical purpose of the present invention, the technical solution adopted by the present invention is as follows: A method for extracting adipoyl-7-ADCA comprises: obtaining adipoyl-7-ADCA fermentation broth filtrate; The adipoyl-7-ADCA fermentation broth filtrate is mixed and stirred with a pre-configured mixed solvent and a surfactant to obtain an initial mixed solution; Adjusting the pH value of the initial mixed solution to obtain a secondary treated mixed solution; and stirring the secondary treated mixed solution at normal pressure to cause the secondary treated mixed solution to reach a turbulent state; The secondary treatment mixed liquid in the turbulent state is extracted and separated to obtain a light phase adipoyl-7-ADCA extract, and the heavy phase waste acid water (raffinate) after separation is recycled in an environmentally friendly manner.
[0007] Optionally, obtaining adipoyl-7-ADCA fermentation broth filtrate comprises: The pre-configured adipoyl-7-ADCA strain is cultured in a seed tank to obtain a seed tank seed liquid; the seed tank seed liquid is fermented to obtain an adipoyl-7-ADCA fermentation liquid; the adipoyl-7-ADCA fermentation liquid is pressure-filtered using a pressure plate frame to obtain an adipoyl-7-ADCA fermentation liquid filtrate.
[0008] Optionally, the mixed solvent includes a mixture of 3-8 carbon alcohols and 3-8 carbon esters, and the volume ratio of the carbon alcohol to the carbon ester is 1:2-3:2.
[0009] Optionally, the volume ratio of the adipoyl-7-ADCA fermentation broth filtrate to the mixed solvent is 1:1 to 3:1.
[0010] Optionally, the total solvent content of the raffinate phase is less than 4%, and the water content of the extraction solvent phase is 7% to 10%, wherein the total solvent content of the raffinate phase refers to the amount of solvent contained in the heavy phase after the filtrate is extracted. That is, the number of grams of solvent contained in 100 ml of the raffinate phase, and the water content of the extraction solvent phase refers to the amount of water contained in the solvent phase after extraction, that is, the number of grams of water contained in 100 ml of the extraction solvent phase.
[0011] Optionally, the surfactant is a pre-configured polyethylene polyamine non-ionic surfactant, and the added amount of the surfactant is 2000-4000 ppm based on the volume of the adipoyl-7-ADCA fermentation broth filtrate.
[0012] Optionally, polyethylene polyamine nonionic surfactants include ethylenediamine, diethylenetriamine, triethylenetetramine, and tetraethylenepentamine; the molecular chains of polyethylene polyamine nonionic surfactants simultaneously have amino groups, polyoxyethylene, organic silicone oxygen, multi-arm and other structures, and their excellent properties are combined to effectively reduce the oil-water interfacial tension, quickly and effectively destroy the stability of the emulsion, and greatly improve the oil-water separation efficiency.
[0013] Optionally, the configuration concentration of the polyethylene polyamine nonionic surfactant is 1%~5%, and the configuration method of the polyethylene polyamine nonionic surfactant includes: adding drinking water to a preparation tank, controlling the temperature at 10℃-25℃, starting stirring, weighing the polyethylene polyamine nonionic surfactant at 1%~5% of the weight of the drinking water, slowly adding the polyethylene polyamine nonionic surfactant to the drinking water, and continuing stirring for 20 minutes.
[0014] Optionally, adjusting the pH value of the initial mixed solution includes: slowly adding a pre-configured dilute sulfuric acid solution to the initial mixed solution for multiple times to adjust the pH value of the initial mixed solution.
[0015] Optionally, the concentration of the dilute sulfuric acid solution is 10% to 30%, preferably, the concentration of the dilute sulfuric acid solution is 20%; and the pH value of the initial mixed solution after pH adjustment is 1.0 to 3.0, preferably, the pH value of the initial mixed solution after pH adjustment is 1.8.
[0016] Optionally, the preparation method of the dilute sulfuric acid solution includes: adding drinking water to a preparation tank, controlling the temperature at 10°C-25°C, starting stirring, calculating the amount of concentrated sulfuric acid to be added based on the dilute sulfuric acid concentration of 10%-30% (the concentrated sulfuric acid is industrial concentrated sulfuric acid (93%)), slowly adding the concentrated sulfuric acid to the drinking water, and continuing stirring for 20 minutes.
[0017] Optionally, the Reynolds number Re of the fluid in the turbulent state is greater than or equal to 4000; and the temperatures of the normal pressure stirring and extraction treatments are both 15-25°C; and the separation treatment is carried out by a centrifuge, and the separation factor Fr of the centrifuge is greater than 2000.
[0018] The technical solution provided by the present invention may include the following beneficial effects: The invention can extract adipoyl-7-ADCA from fermentation broth filtrate into a mixed solvent. The extraction process has high yield, strong impurity removal ability, mild process conditions and is suitable for large-scale production.
[0019] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 The chromatogram of the extraction filtrate before extraction in Example 1; Figure 2 The liquid chromatogram of the extract after extraction in Example 1; Figure 3 The chromatogram of the extraction filtrate before extraction in Example 2; Figure 4 The liquid chromatogram of the extract after extraction in Example 2; Figure 5 The chromatogram of the extraction filtrate before extraction in Example 3; Figure 6 The figure is the liquid chromatogram of the extract after extraction in Example 3. DETAILED DESCRIPTION
[0021] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention belong to the scope of protection of the present invention.
[0022] In the adipoyl-7-ADCA fermentation broth, the active ingredient is adipoyl-7-ADCA salt, which is acid under acidic conditions and easily soluble in organic solvents. It is salt under neutral conditions and easily soluble in water. Adipoyl-7-ADCA salt is relatively stable under weak acid and neutral conditions, but is easily destroyed under strong acid and alkaline conditions. The structural formula of adipoyl-7-ADCA is as follows:
[0023] Based on the above situation, the present invention provides a method for extracting adipoyl-7-ADCA, as shown in the following examples.
[0024] Example 1 A method for extracting adipoyl-7-ADCA according to Example 1 of the present invention comprises the following steps: Add 1730L of adipoyl-7-ADCA fermentation broth filtrate and 860L of mixed solvent (wherein the volume of n-butanol: the volume of butyl acetate is 3:2) into the mixed extraction tank, the extraction phase ratio O / A=1:2.01, and add 140L of 3% concentration of polyethylene polyamine surfactant (the volume of surfactant is 2427ppm of the volume of the filtrate) for demulsification; Then, start the mixing extraction tank to stir, adjust the stirring speed, make the material in the tank reach a turbulent state (Reynolds number Re ≥ 4000), slowly add 22% concentration of dilute sulfuric acid solution, and continuously detect the pH value of the material until the pH value of the material is 1.82, stop adding acid, and the extraction is completed; Prepare for centrifugal separation, pump the mixed liquid after mixed extraction into the centrifuge for separation, the centrifuge separation factor Fr=3000, the feed rate is controlled at 3~4m3 / h, adjust the centrifuge back pressure to ensure that the light phase extract is just clarified, the heavy phase raffinate after separation enters the wastewater tank for environmentally friendly recycling, and the light phase extract enters the product storage tank for further processing.
[0025] The fermentation broth was filtered through a plate and frame to obtain adipoyl-7-ADCA filtrate, with adipoyl-7-ADCA content of 26607ug / ml and pH=6.25. The heavy phase raffinate had a volume of 1485L, adipoyl-7-ADCA content of 3588ug / ml, pH=1.86, butyl acetate content of 0.25%, and n-butanol content of 4.7%. The light phase extract had a volume of 825L, adipoyl-7-ADCA content of 71043ug / ml, and a water content of 9.3%. The distribution coefficient of the extracted adipoyl-7-ADCA content and the heavy phase raffinate volume was: 3588:71043=1:19.8; the extraction ratio was: (71043*825) / (3588*1485)=11:1, and the yield was 91.6%. The filtrate before extraction contained adipoyl-7-ADCA with a purity of 78.18%, and the purity of the extract after extraction was 97.54%. The purity of adipoyl-7-ADCA before and after extraction was shown in the liquid chromatography. Figure 1 and Figure 2 As shown. It can be seen that the extraction process has high yield and strong impurity removal ability.
[0026] Example 2 A method for extracting adipoyl-7-ADCA according to Example 2 of the present invention comprises the following steps: 1850L of adipoyl-7-ADCA fermentation broth filtrate and 800L of mixed solvent (wherein the volume of n-butanol: the volume of butyl acetate is 3:2) were added to the mixed extraction tank, the extraction phase ratio O / A was 1:2.31, and 150L of 3% concentration of polyethylene polyamine surfactant (the volume of surfactant was 2432ppm of the volume of the filtrate) was added for demulsification; Then start the mixing extraction tank to stir, adjust the stirring speed to make the material in the tank reach a turbulent state (Reynolds number Re ≥ 4000), slowly add 22% concentration of dilute sulfuric acid solution, and continuously detect the pH value of the material until the pH value of the material is 1.80, stop adding acid, and the extraction is completed; Prepare for centrifugal separation, pump the mixed liquid after mixed extraction into the centrifuge for separation, the centrifuge separation factor Fr=3000, the feed rate is controlled at 3~4m3 / h, adjust the centrifuge back pressure to ensure that the light phase extract is just clarified, the heavy phase raffinate after separation enters the wastewater tank for environmentally friendly recycling, and the light phase extract enters the product storage tank for further processing.
[0027] The fermentation broth was filtered through a plate and frame to obtain a filtrate of adipoyl-7-ADCA, with a content of adipoyl-7-ADCA of 22630ug / ml and a pH of 6.34. The volume of the heavy phase raffinate was 1440L, the content of adipoyl-7-ADCA was 4765ug / ml, the pH was 1.82, the content of butyl acetate was 0.33%, and the content of n-butanol was 8.3%. The volume of the light phase extract was 720L, the content of adipoyl-7-ADCA was 90534ug / ml, and the water content was 8.7%. The distribution coefficient of the extracted adipoyl-7-ADCA content and the volume of the heavy phase raffinate was: 4765:90534=1:19; the extraction ratio was: (90534*720) / (4765*1440)=9.5:1, and the yield was 90.48%. The purity of adipoyl-7-ADCA in the filtrate before extraction was 77.65%, and the purity of the extract after extraction was 97.46%. The purity of adipoyl-7-ADCA before and after extraction was shown in the liquid chromatography. Figure 3 and Figure 4 As shown. It can be seen that the extraction process has high yield and strong impurity removal ability.
[0028] Example 3 A method for extracting adipoyl-7-ADCA according to Example 3 of the present invention comprises the following steps: Add 1700L of adipoyl-7-ADCA fermentation broth filtrate and 850L of mixed solvent (wherein the volume of n-butanol: the volume of butyl acetate is 3:2) into the mixed extraction tank, the extraction phase ratio O / A=1:2, and add 150L of 3% concentration of polyethylene polyamine surfactant (the volume of surfactant is 2647ppm of the volume of the filtrate) for demulsification; Then start the mixing extraction tank to stir, adjust the stirring speed to make the material in the tank reach a turbulent state (Reynolds number Re ≥ 4000), slowly add 22% concentration of dilute sulfuric acid solution, and continuously detect the pH value of the material until the pH value of the material is 1.69, stop adding acid, and the extraction is completed; Prepare for centrifugal separation, pump the mixed liquid after mixed extraction into the centrifuge for separation, the centrifuge separation factor Fr=3000, the feed rate is controlled at 4~5m3 / h, adjust the centrifuge back pressure to ensure that the light phase extract is just clarified, the heavy phase raffinate after separation enters the wastewater tank for environmentally friendly recycling, and the light phase extract enters the product storage tank for further processing.
[0029] The fermentation broth was filtered through a plate and frame to obtain a filtrate of adipoyl-7-ADCA, with a content of adipoyl-7-ADCA of 27459ug / ml and a pH of 6.35. The volume of the heavy phase raffinate was 1493L, the content of adipoyl-7-ADCA was 3527ug / ml, the pH was 1.61, the content of butyl acetate was 0.21%, and the content of n-butanol was 3.2%. The volume of the light phase extract was 830L, the content of adipoyl-7-ADCA was 72320ug / ml, and the water content was 8.7%. The distribution coefficient of the extracted adipoyl-7-ADCA content and the volume of the heavy phase raffinate was: 3527:72320=1:20.5; the extraction ratio was: (72320*830) / (3527*1493)=11.4:1, and the yield was 91.94%. The purity of adipoyl-7-ADCA in the filtrate before extraction was 77.81%, and the purity of the extract after extraction was 96.47%. The purity of adipoyl-7-ADCA before and after extraction was shown in the liquid chromatography. Figure 5 , Figure 6 As shown. It can be seen that the extraction process has high yield and strong impurity removal ability.
[0030] The present invention is not limited to the structures which have been described above and shown in the drawings, and various modifications and changes may be made without departing from the scope thereof. The scope of the present invention is limited only by the appended claims.
Claims
1. A method for extracting adipoyl-7-ADCA, characterized in that: include: obtaining adipoyl-7-ADCA fermentation broth filtrate; The adipoyl-7-ADCA fermentation broth filtrate is mixed and stirred with a pre-configured mixed solvent and a surfactant to obtain an initial mixed solution; Adjusting the pH value of the initial mixed solution to obtain a secondary treated mixed solution; and stirring the secondary treated mixed solution at normal pressure to cause the secondary treated mixed solution to reach a turbulent state; The turbulent secondary treatment mixed liquid is subjected to extraction and separation treatment to obtain a light phase adipoyl-7-ADCA extract.
2. The method for extracting adipoyl-7-ADCA according to claim 1, wherein Obtaining adipoyl-7-ADCA fermentation broth filtrate includes: The pre-configured adipoyl-7-ADCA strain is cultured in a seed tank to obtain a seed tank seed solution; and the seed tank seed solution is fermented to obtain an adipoyl-7-ADCA fermentation solution; The adipoyl-7-ADCA fermentation broth is pressure filtered by using a pressure plate frame to obtain adipoyl-7-ADCA fermentation broth filtrate.
3. The method for extracting adipoyl-7-ADCA according to claim 1, wherein The mixed solvent includes a mixture of 3-8 carbon alcohols and 3-8 carbon esters, and the volume ratio of the carbon alcohol to the carbon ester is 1:2-3:
2.
4. The method for extracting adipoyl-7-ADCA according to claim 1, wherein The volume ratio of the adipoyl-7-ADCA fermentation broth filtrate to the mixed solvent is 1:1-3:
1.
5. The method for extracting adipoyl-7-ADCA according to claim 1, characterized in that The surfactant is a pre-configured polyethylene polyamine non-ionic surfactant, and the added amount of the surfactant is 2000-4000 ppm of the volume of the adipoyl-7-ADCA fermentation broth filtrate.
6. The method for extracting adipoyl-7-ADCA according to claim 5, characterized in that: The configuration concentration of the polyethylene polyamine nonionic surfactant is 1% to 5%, and the configuration method of the polyethylene polyamine nonionic surfactant includes: Add drinking water to the preparation tank, control the temperature at 10℃-25℃, start stirring, weigh 1%~5% of the weight of drinking water polyethylene polyamine nonionic surfactant, slowly add the polyethylene polyamine nonionic surfactant to the drinking water, and continue stirring for 20 minutes.
7. The method for extracting adipoyl-7-ADCA according to claim 1, characterized in that Adjusting the pH value of the initial mixed solution comprises: The pre-configured dilute sulfuric acid solution is slowly added to the initial mixed solution for multiple times to adjust the pH value of the initial mixed solution.
8. The method for extracting adipoyl-7-ADCA according to claim 7, characterized in that: The concentration of the dilute sulfuric acid solution is 10% to 30%, and the pH value of the initial mixed solution after pH adjustment is 1.0 to 3.
0.
9. The method for extracting adipoyl-7-ADCA according to claim 8, characterized in that: The preparation method of the dilute sulfuric acid solution comprises: Add drinking water to the preparation tank, control the temperature at 10℃-25℃, start stirring, calculate the amount of concentrated sulfuric acid to be added based on the dilute sulfuric acid concentration of 10%~30%, slowly add the concentrated sulfuric acid to the drinking water, and continue stirring for 20 minutes.
10. The method for extracting adipoyl-7-ADCA according to claim 1, characterized in that: The Reynolds number Re of the fluid in the turbulent state is greater than or equal to 4000; and the temperature of the normal pressure stirring and extraction treatment is 15-25°C; the separation treatment is carried out by a centrifuge, and the separation factor Fr of the centrifuge is greater than 2000.