CN108147976B - Extraction method of demethylated aureomycin - Google Patents
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Abstract
The invention provides a method for extracting demethylated aureomycin, which comprises the following steps: adjusting the pH value of the fermentation liquid to acidity, and performing solid-liquid separation on the fermentation liquid and the thalli to obtain liquid 1; carrying out complexation reaction on the obtained liquid 1 and 3-4% of cationic surfactant at the pH value of 8.0-8.5, and carrying out solid-liquid separation to obtain a complex 1; the cationic surfactant is tetradecyl trimethyl ammonium chloride and/or hexadecyl trimethyl ammonium chloride; the percentage is the mass percentage of the cationic surfactant in the mass of the liquid 1; adjusting the pH value of the obtained complex 1, carrying out a complex reaction with urea, and carrying out solid-liquid separation to obtain a complex 2; adjusting the pH value of the obtained complex 2 to dissolve the complex, and performing solid-liquid separation to obtain liquid 2; and (3) regulating the pH value of the obtained liquid 2 by using hydrochloric acid, cooling and crystallizing to obtain the DMCTC hydrochloride. The extraction method of the methyl aureomycin can obtain the DMCTC hydrochloride with the purity of more than or equal to 94 percent.
Description
Technical Field
The invention relates to a method for extracting demethylated aureomycin.
Background
Demethylated aureomycin (abbreviated as DMCTC in English) has a wide antibacterial spectrum, has a strong effect on gram-positive bacteria including tetracycline-resistant staphylococcus aureus, streptococcus and the like and gonorrhea and sirtuin bacteria in gram-negative bacteria, is a key precursor for producing milocycline and tigecycline, and is an important medical raw material.
Demethylated aureomycin is produced by fermenting Streptomyces aureus, and then raw material demethylated aureomycin salt is obtained by separation and extraction, wherein the separation and extraction processes mainly reported at present mainly comprise: (1) the fermentation liquor is subjected to acidification, extraction, back extraction and concentration crystallization (China J.antibiotics 2000, 25: 341-; (2) the demethylated aureomycin (China J.antibiotics, 2013, 38: 915-; (3) adjusting the pH of the fermented liquid to alkaline, and adding seed crystals for crystallization (CN 201310575459). Although the operation is simple, the demethylated aureomycin has poor stability under alkaline conditions, so that the DMCTC epimer is easily formed; (4) adjusting pH of the concentrated decolorized fermentation broth to 4.5-8.5, crystallizing, precipitating, and refining with demethyl aureomycin double salt method (CN 201310380480). But the separation process needs to utilize resin to decolor the fermentation liquor to reduce impurities, thereby increasing the operation cost.
The price of the DMCTC is low, the dosage is large, the application has higher requirement on the purity of the DMCTC, and the purity of the DMCTC prepared by the existing process can not reach more than 94 percent, so that the large-scale production and application of the DMCTC can be limited, and the problem to be solved at present is urgent to develop a simple and feasible production process with low pollution and low cost so as to ensure that the purity of the DMCTC reaches more than 94 percent and even reaches the quality requirement of USP (the purity is more than or equal to 95 percent).
Disclosure of Invention
The invention aims to overcome the defects of large usage amount of organic solvents, high production cost and the like in the prior art, and provides a method for extracting demethylated aureomycin with small pollution and low cost.
The invention provides a method for extracting demethylated aureomycin, which comprises the following steps:
(1) adjusting the pH value of fermentation liquor containing the demethylated aureomycin to be acidic, and carrying out solid-liquid separation on the fermentation liquor and thalli to obtain liquid 1;
(2) carrying out complexation reaction on the obtained liquid 1 and 3-4% of cationic surfactant at the pH value of 8.0-8.5, and carrying out solid-liquid separation to obtain a complex 1; the cationic surfactant is tetradecyl trimethyl ammonium chloride and/or hexadecyl trimethyl ammonium chloride; the percentage is the percentage of the mass of the cationic surfactant to the mass of the liquid 1;
(3) adjusting the pH value of the obtained complex 1, carrying out a complex reaction with urea, and carrying out solid-liquid separation to obtain a complex 2;
(4) adjusting the pH value of the obtained complex 2, dissolving the obtained complex 2, and carrying out solid-liquid separation to obtain liquid 2;
(5) and (3) regulating the pH value of the obtained liquid 2 to be acidic by using hydrochloric acid, and cooling and crystallizing to obtain the DMCTC hydrochloride.
In the invention, the fermentation liquor containing the demethylated aureomycin is fermentation liquor which is produced by fermenting Streptomyces aureus (Streptomyces aureus) and contains the demethylated aureomycin, which is generally known in the field.
In step (1), the operation of adjusting the pH value of the fermentation solution is conventional in the art, the pH value is preferably adjusted to 1.0-2.0, and the pH value is more preferably adjusted to 1.5. The pH adjusting substance is a pH adjusting agent conventional in the art, such as an aqueous sulfuric acid solution.
In step (1), the solid-liquid separation is conventional in the art, and is preferably microfiltration, and the microfiltration membrane used for microfiltration is conventional in the art, and is preferably a 0.45 μm ceramic membrane.
In step (2), the tetradecyltrimethylammonium chloride is conventional in the art, preferably purumtac 150, the hexadecyltrimethylammonium chloride is conventional in the art, preferably purumtac 150, and the purumtac 150 and purumtac 150 are available from korean idchem ltd.
In step (2), the pH adjusting substance is a pH regulator conventional in the art, such as an aqueous solution of sodium hydroxide.
In step (2), the solid-liquid separation is conventional in the art and is typically filtration.
In step (3), the operation of adjusting the pH of the obtained complex 1 is conventional in the art, and the pH is preferably adjusted to 1.0-2.0, more preferably to 1.5. The pH adjusting substance is a pH adjusting agent conventional in the art, such as an aqueous sulfuric acid solution.
In step (3), the solid-liquid separation is conventional in the art and is typically filtration.
In step (4), the operation of adjusting the pH of the obtained complex 2 is conventional in the art, and the pH is preferably adjusted to 2.0-3.0, and more preferably to 2.5. The pH adjusting substance is a pH adjusting agent conventional in the art, such as an aqueous solution of sodium hydroxide.
In step (4), the solid-liquid separation is conventional in the art and is typically filtration.
In step (5), the operation of adjusting the pH of the obtained liquid 2 is conventional in the art, and the pH is preferably adjusted to 0.2 to 0.8, and more preferably to 0.5.
In the step (5), the operation of temperature reduction is conventional in the field, and the temperature reduction is preferably carried out to 5 ℃; the operation of the crystallization is conventional in the art, and the time for the crystallization is preferably 6 hours.
On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows: the extraction method of demethylated aureomycin of the invention can obtain the DMCTC hydrochloride with the purity of more than or equal to 94 percent by creatively discovering the type and the dosage of the cationic surfactant and the pH value after the cationic surfactant is added, and even reaches the USP quality requirement (the purity of more than or equal to 95 percent).
Drawings
FIG. 1 is an HPLC chromatogram of a demethylated aureomycin broth according to example 1 of the present invention.
FIG. 2 is an HPLC chromatogram of demethylchlortetracycline hydrochloride obtained in example 1 of the present invention.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
S. aureus (Streptomyces aureus) ATCC 21658 was purchased from American Type Culture Collection (ATCC).
The fermentation broth containing demethylated aureomycin of the following examples was prepared as follows: inoculating hypha of Streptomyces aureus ATCC 21658 to a seed culture medium, culturing at 28 ℃ for 40 hours, inoculating to a fermentation culture medium according to the inoculation amount of 10%, culturing at 28 ℃ for 7-8 days, collecting fermentation liquor containing demethyl aureomycin, and using the fermentation liquor for next separation and purification.
Seed medium (g/L): 2.0 parts of soluble starch, 1.5 parts of glucose, 1.5 parts of soybean cake powder and 0.3 part of sodium chloride;
fermentation medium (g/L) comprises soluble starch 5.0, maltodextrin 2.0, glucose 1.0, soybean cake powder 3.0, dried yeast powder 5.0, soybean oil 2.0, CaCO3 0.3。
The cationic surfactants PurumtTAC150 and PurumCTAC150 in the following examples were purchased from Korea idchem Co., Ltd.
DMCTC HPLC the analytical method is as follows:
and (3) analysis and column fixing: ODSC18 column, 4.6 × 250mm, 5 um; column temperature: 40 ℃;
mobile phase a was 0.02mol/L potassium dihydrogen phosphate (0.1% phosphoric acid) in water acetonitrile 90: 10(V/V), 0.02mol/L potassium dihydrogen phosphate (0.1% phosphoric acid) water solution of acetonitrile 70: 30 (V/V); the flow rate is 1.0 mL/min; detector wavelength 254 nm; the sample amount is 10uL, and the running time is 20 min.
Gradient elution table:
| time (min) | Phase A (%) | Phase B (%) |
| 0 | 100 | 0 |
| 5 | 100 | 0 |
| 8 | 0 | 100 |
| 18 | 0 | 100 |
| 23 | 100 | 0 |
| 30 | 100 | 0 |
Example 1
The spectrum of the fermentation liquor containing the demethylated aureomycin is shown in figure 1, the fermentation unit is 3250mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 3.0% PurumtTAC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. Adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 deg.C, crystallizing for 6 hr, filtering, collecting filter cake, and drying at 60 deg.C for 8 hr to obtain DMCTC hydrochloride with a spectrum shown in FIG. 2, peak appearance after about 13min, total yield of 74%, and purity of 95.5%.
Example 2
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3250mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 3.5% PurumtTAC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 74.5%, and the purity is 95.4%.
Example 3
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3250mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 4.0% PurumtTAC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 75.1%, and the purity is 95.2%.
Comparative example 1
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3250mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 2.5% PurumtTAC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 63% and the purity is 93.2%.
Comparative example 2
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3250mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 4.5% PurumtTAC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 68% and the purity is 94.3%.
| Item | Overall yield/percentage of DMCTC | Purity/% of DMCTC |
| Example 1 | 74 | 95.5 |
| Example 2 | 74.5 | 95.4 |
| Example 3 | 75.1 | 95.2 |
| Comparative example 1 | 63 | 93.2 |
| Comparative example 2 | 68 | 94.3 |
Example 4
Fermenting the fermentation liquor containing the demethylated aureomycin with the fermentation unit of 3200mg/L and the volume of 1000mL, adjusting the pH value to 1.5 by using a sulfuric acid aqueous solution, stirring for 30 minutes, filtering by using a 0.45 mu m ceramic membrane, and collecting the filtrate. Adding 3.0% PurumCTAC150, then adjusting the pH value to 8.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 60.2%, and the purity is 94.2%.
Example 5
Fermenting the fermentation liquor containing the demethylated aureomycin with the fermentation unit of 3200mg/L and the volume of 1000mL, adjusting the pH value to 1.5 by using a sulfuric acid aqueous solution, stirring for 30 minutes, filtering by using a 0.45 mu m ceramic membrane, and collecting the filtrate. Adding 3.5% PurumCTAC150, then adjusting the pH value to 8.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 63% and the purity is 94.6%.
Example 6
Fermenting the fermentation liquor containing the demethylated aureomycin with the fermentation unit of 3200mg/L and the volume of 1000mL, adjusting the pH value to 1.5 by using a sulfuric acid aqueous solution, stirring for 30 minutes, filtering by using a 0.45 mu m ceramic membrane, and collecting the filtrate. Adding 4.0% PurumCTAC150, then adjusting the pH value to 8.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 62%, and the purity is 94.2%.
Comparative example 3
Fermenting the fermentation liquor containing the demethylated aureomycin with the fermentation unit of 3200mg/L and the volume of 1000mL, adjusting the pH value to 1.5 by using a sulfuric acid aqueous solution, stirring for 30 minutes, filtering by using a 0.45 mu m ceramic membrane, and collecting the filtrate. Adding 2.5% PurumCTAC150, then adjusting the pH value to 8.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 59 percent, and the purity is 91.1 percent.
Comparative example 4
Fermenting the fermentation liquor containing the demethylated aureomycin with the fermentation unit of 3200mg/L and the volume of 1000mL, adjusting the pH value to 1.5 by using a sulfuric acid aqueous solution, stirring for 30 minutes, filtering by using a 0.45 mu m ceramic membrane, and collecting the filtrate. Adding 4.5% PurumCTAC150, then adjusting the pH value to 8.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 60 percent, and the purity is 92.8 percent.
| Item | Overall yield/percentage of DMCTC | Purity/% of DMCTC |
| Example 4 | 60.2 | 94.2 |
| Example 5 | 63 | 94.6 |
| Example 6 | 62 | 94.2 |
| Comparative example 3 | 59 | 91.1 |
| Comparative example 4 | 60 | 92.8 |
Example 7
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3500mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 3.0% PurumtTAC150 was added, then the pH was adjusted to 8.0 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 73 percent, and the purity is 95.8 percent.
Example 8
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3500mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 3.0% PurumtTAC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 74 percent, and the purity is 95.5 percent.
Comparative example 5
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3500mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 3.0% PurumtTAC150 was added, then the pH was adjusted to 7.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 40% and the purity is 93.3%.
Comparative example 6
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3500mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. 3.0% PurumtTAC150 was added, then the pH was adjusted to 9.0 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 61 percent, and the purity is 92.4 percent.
| Item | Overall yield/percentage of DMCTC | Purity/% of DMCTC |
| Example 7 | 73 | 95.8 |
| Example 8 | 74 | 95.5 |
| Comparative example 5 | 40 | 93.3 |
| Comparative example 6 | 61 | 92.4 |
Example 9
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3230mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. Adding 3.0% PurumCTAC150, then adjusting the pH value to 8.0 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 58% and the purity is 94.2%.
Example 10
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3230mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. Adding 3.0% PurumCTAC150, then adjusting the pH value to 8.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 60%, and the purity is 94.2%.
Comparative example 7
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3230mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. Adding 3.0% PurumCTAC150, then adjusting the pH value to 7.5 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 52% and the purity is 93.7%.
Comparative example 8
The fermentation unit of the fermentation liquor containing the demethylated aureomycin is 3230mg/L, the volume is 1000mL, the pH value is adjusted to 1.5 by using a sulfuric acid aqueous solution, the mixture is stirred for 30 minutes, and the filtrate is collected after being filtered by a 0.45 mu m ceramic membrane. Adding 3.0% PurumCTAC150, then adjusting the pH value to 9.0 with NaOH aqueous solution, filtering with a plate frame, and collecting a filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. And adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, cooling to 5 ℃, crystallizing for 6 hours, filtering, collecting a filter cake, and drying at 60 ℃ for 8 hours to obtain the DMCTC hydrochloride, wherein the total yield is 59 percent, and the purity is 93.5 percent.
| Item | Overall yield/percentage of DMCTC | Purity/% of DMCTC |
| Example 9 | 58 | 94.2 |
| Example 10 | 60 | 94.2 |
| Comparative example 7 | 52 | 93.7 |
| Comparative example 8 | 59 | 93.5 |
Comparative example 9 fermentation broth containing demethylated aureomycin, having a fermentation unit of 3200mg/L and a volume of 1000mL, was adjusted to pH 1.5 with aqueous sulfuric acid, stirred for 30 minutes, filtered through a 0.45 μm ceramic membrane and the filtrate was collected. Adding 4.0% PurumCECP, adjusting pH to 8.5 with NaOH water solution, filtering with plate frame, and collecting filter cake. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. Adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, and cooling to 5 ℃. Crystallizing for 6 hours, filtering, collecting a filter cake, and drying for 8 hours at 60 ℃ to obtain the DMCTC hydrochloride.
Comparative example 10 fermentation broth containing demethylated aureomycin, having a fermentation unit of 3200mg/L and a volume of 1000mL, was adjusted to pH 1.5 with aqueous sulfuric acid, stirred for 30 minutes, filtered through a 0.45 μm ceramic membrane and the filtrate was collected. 4.0% PurumBC150 was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtration was performed, and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. Adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, and cooling to 5 ℃. Crystallizing for 6 hours, filtering, collecting a filter cake, and drying for 8 hours at 60 ℃ to obtain the DMCTC hydrochloride.
Comparative example 11 fermentation broth containing demethylated aureomycin, having a fermentation unit of 3200mg/L and a volume of 1000mL, was adjusted to pH 1.5 with aqueous sulfuric acid, stirred for 30 minutes, filtered through a 0.45 μm ceramic membrane and the filtrate was collected. 4.0% PurumBTAC was added, then the pH was adjusted to 8.5 with aqueous NaOH, plate and frame filtered and the filter cake was collected. After washing with water, the pH value was adjusted to 1.5 with an aqueous solution of sulfuric acid, urea was added while maintaining the pH value at 1.5, and the filter cake was collected by filtration. Adjusting pH to 2.5 with NaOH water solution, stirring, filtering, and collecting filtrate. Adding hydrochloric acid into the filtrate until the pH value is 0.5, stirring, and cooling to 5 ℃. Crystallizing for 6 hours, filtering, collecting a filter cake, and drying for 8 hours at 60 ℃ to obtain the DMCTC hydrochloride.
| Item | Overall yield/percentage of DMCTC | Purity/% of DMCTC |
| Comparative example 9 | 20 | 90.5 |
| Comparative example 10 | 35 | 89.8 |
| Comparative example 11 | 38 | 86.2 |
Comparative example 12
The inventors carried out the following experiments according to the method described in patent CN201310575459, and obtained purity data.
Fermenting the fermentation liquor containing the demethylated aureomycin with a fermentation unit of 3200mg/L and a volume of 1000ml, adjusting the pH value to 1.5 by using a sulfuric acid aqueous solution, stirring for 30 minutes, filtering by using a 0.45 mu m ceramic membrane, and collecting the filtrate. Cooling to 8 ℃, adjusting the pH value to 9.2 by using NaOH aqueous solution, separating out crystals, filtering by using a plate frame, and collecting filter cakes. After being washed by water, the pH value is adjusted to 2.8 by hydrochloric acid, and the product is dried in vacuum, so that 1.7g of the product is obtained, the total yield is 53.1 percent, and the purity is 84.5 percent.
Claims (8)
1. A method for extracting demethylated aureomycin is characterized in that: which comprises the following steps:
(1) adjusting the pH value of fermentation liquor containing the demethylated aureomycin to be acidic, and carrying out solid-liquid separation on the fermentation liquor and thalli to obtain liquid 1;
in the step (1), the pH value is adjusted to 1.0-2.0;
(2) carrying out complexation reaction on the obtained liquid 1 and 3-4% of cationic surfactant at the pH value of 8.0-8.5, and carrying out solid-liquid separation to obtain a complex 1; the cationic surfactant is tetradecyl trimethyl ammonium chloride and/or hexadecyl trimethyl ammonium chloride; the percentage is the percentage of the mass of the cationic surfactant to the mass of the liquid 1;
(3) adjusting the pH value of the obtained complex 1, carrying out a complex reaction with urea, and carrying out solid-liquid separation to obtain a complex 2;
in the step (3), the pH value is adjusted to 1.0-2.0;
(4) adjusting the pH value of the obtained complex 2, dissolving the obtained complex 2, and filtering to obtain liquid 2;
in the step (4), the pH value is adjusted to 2.0-3.0;
(5) regulating the pH value of the obtained liquid 2 with hydrochloric acid, cooling and crystallizing to obtain demethyl aureomycin hydrochloride;
in the step (5), the pH value is adjusted to 0.2-0.8.
2. The extraction method according to claim 1, characterized in that: in the step (1), the pH value is adjusted to 1.5.
3. The extraction method according to claim 1, characterized in that: in the step (1), the solid-liquid separation is microfiltration.
4. The extraction method according to claim 3, characterized in that: the microfiltration membrane adopted by the microfiltration is a 0.45 mu m ceramic membrane.
5. The extraction method according to claim 1, characterized in that: in the step (3), the pH value is adjusted to 1.5.
6. The extraction method according to claim 1, characterized in that: in the step (4), the pH value is adjusted to 2.5.
7. The extraction method according to claim 1, characterized in that: in the step (5), the pH value is adjusted to 0.5.
8. The extraction method according to claim 1, characterized in that: in the step (5), the temperature is reduced to 5 ℃; the time for the crystallization was 6 hours.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3053893A (en) * | 1960-07-22 | 1962-09-11 | American Cyanamid Co | Recrystallization of demethylchlortetracycline |
| GB917467A (en) * | 1959-12-09 | 1963-02-06 | American Cyanamid Co | Improvements in or relating to antibiotics |
| CN103641739A (en) * | 2013-11-18 | 2014-03-19 | 宁夏泰瑞制药股份有限公司 | Method for producing demeclocycline hydrochloride by utilizing demeclocycline fermentation broth |
| CN104418767A (en) * | 2013-08-28 | 2015-03-18 | 河南天方药业股份有限公司 | Demeclocycline extraction process |
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2016
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| GB917467A (en) * | 1959-12-09 | 1963-02-06 | American Cyanamid Co | Improvements in or relating to antibiotics |
| US3053893A (en) * | 1960-07-22 | 1962-09-11 | American Cyanamid Co | Recrystallization of demethylchlortetracycline |
| CN104418767A (en) * | 2013-08-28 | 2015-03-18 | 河南天方药业股份有限公司 | Demeclocycline extraction process |
| CN103641739A (en) * | 2013-11-18 | 2014-03-19 | 宁夏泰瑞制药股份有限公司 | Method for producing demeclocycline hydrochloride by utilizing demeclocycline fermentation broth |
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