CN106589075B - Purification method of teicoplanin - Google Patents
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- C07K9/008—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure directly attached to a hetero atom of the saccharide radical, e.g. actaplanin, avoparcin, ristomycin, vancomycin
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Abstract
The invention provides a purification method of teicoplanin, which comprises the steps of sequentially carrying out resin enrichment, resin purification, membrane concentration and spray drying on teicoplanin fermentation liquor. Compared with the prior art, the method has the advantages that the polar macroporous adsorption resin is combined with the medium-polarity chromatographic resin with uniform particle size, the product yield is high, and the decoloring effect is good; the spray drying technology avoids the use of a large amount of acetone, thereby not only protecting the environment and reducing the cost, but also avoiding the problem of organic solvent residue in the prior art; the proportion of each component of teicoplanin obtained by the purification method of the invention meets the EP standard, the purification process is efficient and convenient, the preparation amount is large, and the method is suitable for industrial production.
Description
[ technical field ] A method for producing a semiconductor device
The invention belongs to the field of pharmaceutical engineering, and particularly relates to a purification method of teicoplanin.
[ background of the invention ]
Teicoplanin (teicoplanin) is another glycopeptide antibiotic against drug-resistant bacteria developed after vancomycin. In the existing teicoplanin production method, the production of teicoplanin by microbial fermentation is generally adopted as a main method, and the teicoplanin in the fermentation liquor comprises TAl-TA 2, wherein TA2 is the main active component of teicoplanin and mainly comprises 5 compounds (TA2-1, TA2-2, TA2-3, TA2-4 and TA2-5) with very similar chemical structures; the crude product and refined product also contain an active component TA3-1, which is deacylated glucosamine product of TA 2; in addition, it contains 2 more lipophilic analogs. The chemical structure of teicoplanin is shown in the following formula I. At present, in the standards followed in the pharmaceutical field, the European Pharmacopoeia (EP) makes clear regulations on the individual components, the sum of the A2 groups is not less than 80.0%, wherein the A2-2 is 35.0% -55.0%, the A2-1, the A2-3, the A2-4 and the A2-5 are not more than 20.0%, the A3-1 is not more than 15.0%, and the impure component is not more than 5.0%.
The existing main purification method of teicoplanin comprises the following steps: solvent extraction, adsorption, ion exchange, chromatography, etc. The solvent extraction method has poor separation and purification effects and low yield, and is rarely adopted at present; the influence of ion exchange on the purification of teicoplanin is described in the literature [ see Journal of Antibiotics, volume 31, No. 3, page 170-177 (3 months 1978) ] of m.r. balton (M.R, Bardon), et al, strong acid ion exchange resins decompose part of the sugar components of teicoplanin, and strong base ion exchange resins epimerize teicoplanin and reduce its biological activity; chinese patent with publication number CN101302248A and US patent with publication number US2005245481A1 both disclose the purification of teicoplanin by liquid chromatography, but the purification methods disclosed by these patents cannot control the proportion of each component of A2 well, cannot reach the standard of European pharmacopoeia, and have long purification time and long production period. Chinese patent with application number CN201210221636.X discloses a method for preparing teicoplanin fine powder by utilizing nano polymer microspheres, and the method partially realizes the content control of single components in the product; however, the cost of the nano polymer microspheres is high, and the nano polymer microspheres are difficult to popularize on a large scale. In addition, chinese patent application No. CN200710107185.6 discloses a method for preparing high-purity teicoplanin by chromatography using sephadex or agarose gel from GE company, but since sephadex and agarose gel are expensive, they are not suitable for industrial production.
Therefore, the existing purification method of teicoplanin is mature and common and is also a resin adsorption method; alkalizing the fermentation liquor, filtering, adsorbing the filtrate with resin, desorbing the resin with a desorbent, decolorizing the desorbed solution with active carbon, precipitating the filtrate with a solvent, filtering to obtain a wet teicoplanin product, refining with a resin or a chromatographic agent, wherein the resin is mainly macroporous adsorption resin, the desorbent is an aqueous solution of an organic solvent, and precipitating with a large amount of the solvent.
Although the above resin adsorption method has been well recognized in specific industrial production, it has the following drawbacks: most of the resins adopted by the first current resin adsorption method are nonpolar macroporous adsorption resins, which have great influence on the control of the proportion of each component of teicoplanin and have unsatisfactory decoloration and impurity removal effects; the second is the final precipitation operation, which usually adopts acetone added to reach 90% acetone concentration to separate out teicoplanin, thus needing a large amount of acetone, greatly increasing the pressure of environmental protection and influencing the production environment and the health of personnel.
In view of the above, it is highly desirable for practitioners to develop a resin adsorption method for purifying teicoplanin, which is suitable for industrial production and is environmentally friendly.
[ summary of the invention ]
The invention aims to solve the technical problem of providing a method for purifying teicoplanin, which is not only environment-friendly, but also suitable for industrial production.
The invention solves the technical problems through the following technical scheme: a purification method of teicoplanin specifically comprises the following operation steps:
(1) resin enrichment: taking teicoplanin fermentation liquor, adjusting the pH value of the teicoplanin fermentation liquor to 10.0-12.0, and filtering to obtain teicoplanin filtrate; pumping the teicoplanin filtrate into a pretreated polar macroporous adsorption resin chromatographic column at the rate of 0.2-2 times of the volume of the packed bed per hour, wherein the size of resin particles of the polar macroporous adsorption resin chromatographic column is 20-60 meshes; then, washing the mixture by purified water until the mixture is colorless, performing gradient elution by taking an aqueous solution of a strong polar solvent as an analytic solution at an elution rate of 0.2-2 times the volume of a packed bed per hour, wherein the analytic solution used in each gradient elution is 3-5 times the volume of the packed bed, and collecting the enrichment solution to obtain a teicoplanin enrichment solution;
(2) resin purification: diluting the teicoplanin concentrated solution obtained in the step (1) by using isometric purified water, then passing through a pretreated medium-polarity macroporous adsorption resin column at a rate of 0.2-2 times the volume of a packed bed per hour, washing the solution to be colorless by using purified water, then performing isocratic elution by using a buffered saline solution of a strong polar solvent as an analytical solution at an elution rate of 0.2-2 times the volume of the packed bed per hour, collecting the eluates step by step, combining effective eluates to obtain a combined solution, detecting the component proportion of the obtained combined solution by using a high performance liquid chromatography, and adjusting the pH value of the combined solution to 6.8-7.4 when each component of teicoplanin in the combined solution meets the requirement of EP (environmental protection) by detecting the high performance liquid chromatography;
(3) and (3) membrane concentration: concentrating the combined solution after the pH value is adjusted in the step (2) by adopting a nanofiltration membrane, and adding purified water with the volume of 3-5 times that of the combined solution during concentration to obtain a teicoplanin concentrated solution;
(4) spray drying: pumping the teicoplanin concentrated solution obtained in the step (3) into a constant flow pump at the flow rate of 1-5L/h for spray drying, and obtaining a teicoplanin dry product meeting the EP standard.
Further, in the step (1), the filtration is performed by plate-and-frame filtration.
Further, in the step (1), the resin of the polar macroporous adsorption resin chromatographic column is acrylamide as a matrix.
Further, in the step (1), the resin of the polar macroporous adsorption resin chromatographic column is HT60 resin.
Further, in the step (1), the gradient of gradient elution is a solvent system of strong polar solvent-water in a volume ratio of 10:90, 20:80, 30:70 and 40:60 respectively.
Further, in the step (2), the resin of the medium-polarity macroporous adsorption resin column is polyacrylate-based, and the particle size of the resin is 100 meshes.
Further, in the step (2), the resin of the medium-polarity macroporous adsorption resin column is HN60 resin.
Further, in the step (2), the buffered saline solution of the strong polar solvent is prepared by using the strong polar solvent and the buffered saline solution in a volume ratio of 40:60, and the concentration of the buffered saline solution is 0.05mol/L, pH, which is 2.5.
Further, in the step (1) and the step (2), the strong polar solvent is low polyhydric alcohol; the buffer salt in the buffer salt water solution is one of sodium dihydrogen phosphate or potassium dihydrogen phosphate.
Further, the strong polar solvent is one of methanol or ethanol.
Further, in the step (3), the nanofiltration membrane is a polyamide membrane, and the molecular weight cut-off of the nanofiltration membrane is 100-1000D; the preferred molecular weight cut-off is 500D.
Further, in the step (4), the spray drying is performed by using a spray dryer, and the specific conditions are as follows: the inlet temperature of the spray dryer is 90-100 ℃, and the outlet temperature is 70-80 DEG C
The invention has the beneficial effects that:
by adopting the polar macroporous adsorption resin and the medium-polarity chromatographic resin with uniform particle size for matching use, the obtained product, namely the teicoplanin dry product which meets the EP standard, has high yield and good decoloration effect; the spray drying is combined, so that the use of an organic solvent is avoided, the environment is protected, the cost is reduced, and the problem of organic solvent residue in the prior art is avoided; in addition, the proportion of each component of teicoplanin obtained by the purification method of the invention meets the EP standard, the operation is efficient and convenient, the preparation amount is large, and the method is suitable for large-scale production.
[ description of the drawings ]
The invention will be further described with reference to the following examples with reference to the accompanying drawings.
FIG. 1 is an HPLC chromatogram of teicoplanin fermentation broth produced by fermentation of Streptomyces according to the present invention.
FIG. 2 is an HPLC chromatogram of enriched solution of HT60 resin teicoplanin in example 1 of the present invention.
FIG. 3 is an HPLC chromatogram of a solution of example 1 of the present invention which is a mixture of teicoplanin and HN60 resin.
FIG. 4 is an HPLC chromatogram of teicoplanin prepared by spray drying according to example 1 of the present invention.
[ detailed description ] embodiments
The invention relates to a purification method of teicoplanin, which comprises the following operation steps:
(1) resin enrichment: taking teicoplanin fermentation liquor, adjusting the pH value of the teicoplanin fermentation liquor to 10.0-12.0, and filtering to obtain teicoplanin filtrate; pumping the teicoplanin filtrate into a pretreated polar macroporous adsorption resin chromatographic column at the rate of 0.2-2 times of the volume of the packed bed per hour, wherein the size of resin particles of the polar macroporous adsorption resin chromatographic column is 20-60 meshes; then, washing the mixture by purified water until the mixture is colorless, performing gradient elution by taking an aqueous solution of a strong polar solvent as an analytic solution at an elution rate of 0.2-2 times the volume of a packed bed per hour, wherein the analytic solution used in each gradient elution is 3-5 times the volume of the packed bed, and collecting the enrichment solution to obtain a teicoplanin enrichment solution;
(2) resin purification: diluting the teicoplanin concentrated solution obtained in the step (1) by using isometric purified water, then passing through a pretreated medium-polarity macroporous adsorption resin column at a rate of 0.2-2 times the volume of a packed bed per hour, washing the solution to be colorless by using purified water, then performing isocratic elution by using a buffered saline solution of a strong polar solvent as an analytic solution at an elution rate of 0.2-2 times the volume of the packed bed per hour, collecting the eluent step by step, combining effective eluents (the eluent containing each component of teicoplanin is the effective eluent) to obtain a combined solution, detecting the component proportion of the obtained combined solution by using a high performance liquid chromatography, and adjusting the pH value of the combined solution to 6.8-7.4 when each component of teicoplanin in the combined solution is determined to meet the EP requirement;
(3) and (3) membrane concentration: concentrating the combined solution after the pH value is adjusted in the step (2) by adopting a nanofiltration membrane, and adding purified water with the volume of 3-5 times that of the combined solution during concentration to obtain a teicoplanin concentrated solution;
(4) spray drying: pumping the teicoplanin concentrated solution obtained in the step (3) into a constant flow pump at the flow rate of 1-5L/h for spray drying, and obtaining a teicoplanin dry product meeting the EP standard.
Wherein, in the step (1), the filtering of the teicoplanin fermentation liquor adopts plate-and-frame filtering; the resin of the polar macroporous adsorption resin chromatographic column is acrylamide-based, such as HT60 resin; the gradient of the gradient elution is a solvent system of strong polar solvent-water with the volume ratio of 10:90, 20:80, 30:70 and 40:60 respectively; the strong polar solvent in the aqueous solution of the strong polar solvent is low-polyhydric alcohol, such as one selected from methanol or ethanol.
In the step (2), the resin of the medium-polarity macroporous adsorption resin column is polyacrylate-based, such as HN60 resin, and the size of the resin particle is 100 meshes; the buffer saline solution of the strong polar solvent is prepared by using the strong polar solvent and the buffer saline solution with the volume ratio of 40:60, wherein the strong polar solvent is low-polyhydric alcohol (such as one of methanol or ethanol), the concentration of the buffer saline solution is 0.05mol/L, pH value and is 2.5, and specifically, the buffer salt in the buffer saline solution is one of sodium dihydrogen phosphate or potassium dihydrogen phosphate.
In the step (3), the nanofiltration membrane is a polyamide membrane, and the molecular weight cut-off of the nanofiltration membrane is 100-1000D; the preferred molecular weight cut-off is 500D. In the step (4), spray drying is carried out by adopting a spray dryer, and the specific conditions are as follows: the inlet temperature of the spray dryer is 90-100 ℃, and the outlet temperature is 70-80 ℃.
It should be noted that the teicoplanin fermentation liquid related in the invention is generated by streptomyces fermentation, and the teicoplanin fermentation liquid is subjected to high performance liquid chromatography detection, and the detection result is shown in fig. 1, so that not only is the proportion of each component of teicoplanin generated by fermentation fully known, but also the comparison and the inspection of the elution effect of the resin used later are facilitated.
For better illustration of the present invention, the present invention is illustrated in the following examples, and for better effect of the treatment, in each example: polar macroporous adsorbent resins and medium polar chromatographic resins are commercially available from Shanghai Min Yongchun; in addition, in order to achieve the purposes of removing impurities in the resin filled in the chromatographic column, activating ions of the resin and the like, the resin needs to be pretreated, and a conventional treatment method is adopted, and the specific operation is as follows: the resin to be used for column filling is repeatedly washed by hot water (clean tap water can also be used) at 50-60 ℃ until the washing water is not brown and has little foam, then the resin treated by the hot water is filled into a chromatographic column, then the resin is treated by acetone with the volume of 5-8 columns, the resin is washed until the effluent liquid is not white and turbid after being added with water, and then the resin column or the chromatographic column is washed by purified water until the acetone smell does not exist, so that the pretreated resin column or the chromatographic column is obtained for later use.
Example 1
Taking 10L of teicoplanin fermentation liquor, adjusting the pH value to 11.0, and filtering by adopting a plate frame to obtain teicoplanin filtrate; the teicoplanin filtrate was pumped through a pretreated polar macroporous adsorption HT60(20-60 mesh) resin chromatography column (Φ 10 × 120cm) at a rate of 1.2 packed bed volumes per hour; then, purifying and washing the mixture until the mixture is colorless, then carrying out gradient elution by using ethanol-water solutions (analytic solutions) with the gradient of volume ratios of 10:90, 20:80, 30:70 and 40:60 in sequence at an elution rate of 1.5L/h, wherein the analytic solution used in each gradient elution is 3 times of the volume of a packed bed, collecting the enrichment solution to obtain teicoplanin enrichment solution, and detecting the teicoplanin enrichment solution by adopting a high performance liquid chromatography, wherein the detection result is shown in figure 2;
diluting the obtained teicoplanin enrichment solution with equal volume of purified water, and passing through a pretreated middle polarity HN60(100 mesh) macroporous adsorption resin column (phi 10 × 120 cm); then, purifying and washing the mixture until the mixture is colorless, performing isocratic elution by using ethanol-sodium dihydrogen phosphate aqueous solution (the concentration of the sodium dihydrogen phosphate aqueous solution is 0.05mol/L, pH 2.5.5) with the volume ratio of 40:60 at the elution rate of 1.5L/h, collecting the eluates step by step, merging effective eluates, detecting the component proportion of the merged solution by adopting high performance liquid chromatography, and adjusting the pH value of the merged solution to 6.8-7.4 when the detection result is shown in figure 3, wherein each component of teicoplanin in the merged solution meets the requirement of EP; concentrating the combined solution after the pH value is adjusted by adopting a polyamide membrane with the molecular weight cutoff of 500D, and adding purified water with the volume of 3-5 times that of the combined solution during concentration to obtain a teicoplanin concentrated solution; and finally, pumping the teicoplanin concentrated solution into a spray dryer for spray drying at the flow rate of 1L/h by a constant flow pump (the conditions of spray drying are that the inlet temperature of the spray dryer is 90-100 ℃, and the outlet temperature is 70-80 ℃) to obtain a teicoplanin dry product, detecting the teicoplanin dry product by adopting a high performance liquid chromatography, wherein the detection result is shown in figure 4, namely the obtained teicoplanin dry product meets the EP standard, weighing is carried out, and 15.7g of the teicoplanin dry product is obtained in the embodiment.
Example 2
Taking 100L of teicoplanin fermentation liquor, adjusting the pH value to 10.0, and filtering by adopting a plate frame to obtain teicoplanin filtrate; the teicoplanin filtrate was pumped through a pretreated polar macroporous adsorption HT60(20-60 mesh) resin chromatography column (Φ 25 × 200cm) at a rate of 0.2 packed bed volumes per hour; then, purifying and washing the mixture until the mixture is colorless, then carrying out gradient elution by using ethanol-water solutions (analytic solutions) with the volume ratios of 10:90, 20:80, 30:70 and 40:60 in sequence at the elution rate of 20L/h, wherein the analytic solution used in each gradient elution is 5 times of the volume of a packed bed, and collecting the enrichment solution to obtain teicoplanin enrichment solution;
diluting the obtained teicoplanin enrichment solution with equal volume of purified water, and passing through a pretreated middle polarity HN60(100 mesh) macroporous adsorbent resin column (phi 25 × 200 cm); then, purifying and washing the mixture until the mixture is colorless, performing isocratic elution by using an ethanol-potassium dihydrogen phosphate aqueous solution (the concentration of the potassium dihydrogen phosphate aqueous solution is 0.05mol/L, pH 2.5.5) with the volume ratio of 40:60 at the elution rate of 15L/h, collecting the eluates step by step, combining the effective eluates, detecting the component proportion of the combined solution by adopting a high performance liquid chromatography, determining that each component of teicoplanin in the combined solution meets the requirement of EP, and adjusting the pH value of the combined solution to 6.8-7.4; concentrating the combined solution after the pH value is adjusted by adopting a polyamide membrane with the molecular weight cutoff of 100D, and adding purified water with the volume of 3-5 times that of the combined solution during concentration to obtain a teicoplanin concentrated solution; finally, the teicoplanin concentrated solution is pumped into a spray dryer for spray drying at the flow rate of 2L/h by a constant flow pump, and the conditions of the spray drying are as follows: the inlet temperature of the spray dryer is 90-100 ℃, and the outlet temperature is 70-80 ℃, so that 185.2g of teicoplanin dry product meeting the EP standard is obtained.
Example 3
Taking 800L of teicoplanin fermentation liquor, adjusting the pH value to 12.0, and then filtering by adopting a plate frame to obtain teicoplanin filtrate; the teicoplanin filtrate was pumped into a pretreated polar macroporous adsorption HT60(20-60 mesh) resin chromatography column (Φ 50 × 300cm) at a rate of 2 packed bed volumes per hour; then, purifying and washing the mixture until the mixture is colorless, then carrying out gradient elution by using ethanol-water solutions (analytic solutions) with the volume ratios of 10:90, 20:80, 30:70 and 40:60 in sequence at the elution rate of 60L/h, wherein the analytic solution used in each gradient elution is 5 times of the volume of a packed bed, and collecting the enrichment solution to obtain teicoplanin enrichment solution;
diluting the obtained teicoplanin enrichment solution with equal volume of purified water, and passing through a pretreated middle polarity HN60(100 mesh) macroporous adsorption resin column (phi 60 × 250 cm); then, purifying and washing the mixture until the mixture is colorless, then, carrying out isocratic elution by using an ethanol-sodium dihydrogen phosphate aqueous solution (the concentration of the sodium dihydrogen phosphate aqueous solution is 0.05mol/L, pH 2.5.5) with the volume ratio of 40:60 at the elution rate of 40L/h, collecting the eluent step by step, combining effective eluents, detecting the component proportion of the combined solution by adopting a high performance liquid chromatography, and adjusting the pH value of the combined solution to 6.8-7.4 when the components of teicoplanin in the combined solution meet the requirement of EP; concentrating the combined solution after the pH value is adjusted by adopting a polyamide membrane with the molecular weight cutoff of 1000D, and adding purified water with the volume of 3-5 times that of the combined solution during concentration to obtain a teicoplanin concentrated solution; finally, the teicoplanin concentrated solution is pumped into a spray dryer for spray drying at the flow rate of 5L/h by a constant flow pump, and the conditions of the spray drying are as follows: the inlet temperature of the spray dryer is 90-100 ℃, and the outlet temperature is 70-80 ℃, so that 486.8g of teicoplanin dry product meeting the EP standard is obtained.
In conclusion, the polar macroporous adsorption resin and the medium-polarity chromatographic resin with uniform particle size are used in a matching manner, so that the obtained product, namely the teicoplanin dry product which meets the EP standard, is high in yield and good in decoloring effect; the spray drying is combined, so that the use of an organic solvent is avoided, the environment is protected, the cost is reduced, and the problem of organic solvent residue in the prior art is avoided; in addition, the proportion of each component of teicoplanin obtained by the purification method of the invention meets the EP standard, the operation is efficient and convenient, the preparation amount is large, and the method is suitable for large-scale production.
Although specific embodiments of the invention have been described above, it will be understood by those skilled in the art that the specific embodiments described are illustrative only and are not limiting upon the scope of the invention, and that equivalent modifications and variations can be made by those skilled in the art without departing from the spirit of the invention, which is to be limited only by the appended claims.
Claims (6)
1. A method for purifying teicoplanin is characterized in that: the purification method specifically comprises the following operation steps:
(1) resin enrichment: taking teicoplanin fermentation liquor, adjusting the pH value of the teicoplanin fermentation liquor to 10.0-12.0, and filtering to obtain teicoplanin filtrate; pumping the teicoplanin filtrate into a pretreated polar macroporous adsorption resin chromatographic column at the rate of 0.2-2 times of the volume of the packed bed per hour, wherein the size of resin particles of the polar macroporous adsorption resin chromatographic column is 20-60 meshes; then, washing the mixture by purified water until the mixture is colorless, performing gradient elution by taking an aqueous solution of a strong polar solvent as an analytic solution at an elution rate of 0.2-2 times the volume of a packed bed per hour, wherein the analytic solution used in each gradient elution is 3-5 times the volume of the packed bed, and collecting the enrichment solution to obtain a teicoplanin enrichment solution;
(2) resin purification: diluting the teicoplanin concentrated solution obtained in the step (1) by using isometric purified water, then passing through a pretreated medium-polarity macroporous adsorption resin column at a rate of 0.2-2 times the volume of a packed bed per hour, washing the solution to be colorless by using purified water, then performing isocratic elution by using a buffered saline solution of a strong polar solvent as an analytical solution at an elution rate of 0.2-2 times the volume of the packed bed per hour, collecting the eluates step by step, combining effective eluates to obtain a combined solution, detecting the component proportion of the obtained combined solution by using a high performance liquid chromatography, and adjusting the pH value of the combined solution to 6.8-7.4 when each component of teicoplanin in the combined solution meets the requirement of EP (environmental protection) by detecting the high performance liquid chromatography;
(3) and (3) membrane concentration: concentrating the combined solution after the pH value is adjusted in the step (2) by adopting a nanofiltration membrane, and adding purified water with the volume of 3-5 times that of the combined solution during concentration to obtain a teicoplanin concentrated solution;
(4) spray drying: pumping the teicoplanin concentrated solution obtained in the step (3) into a constant flow pump at the flow rate of 1-5L/h for spray drying to obtain a teicoplanin dry product meeting the EP standard; the spray drying is carried out by adopting a spray dryer, and the specific conditions are as follows: the inlet temperature of the spray dryer is 90-100 ℃, and the outlet temperature is 70-80 ℃;
in the step (1), the resin of the polar macroporous adsorption resin chromatographic column is HT60 resin;
in the step (2), the resin of the medium-polarity macroporous adsorption resin column is HN60 resin, and the particle size of the resin is 100 meshes;
in the step (3), the nanofiltration membrane is a polyamide membrane, and the molecular weight cut-off of the nanofiltration membrane is 100-1000D.
2. The method for purifying teicoplanin according to claim 1, wherein: in the step (1), the filtration is performed by plate-and-frame filtration.
3. The method for purifying teicoplanin according to claim 1, wherein: in the step (1), gradient elution is sequentially performed by strong polar solvent-water solvent systems with volume ratios of 10:90, 20:80, 30:70 and 40:60 respectively.
4. The method for purifying teicoplanin according to claim 1, wherein: in the step (2), the buffered saline solution of the strong polar solvent is prepared by the strong polar solvent and the buffered saline solution in a volume ratio of 40:60, and the concentration of the buffered saline solution is 0.05mol/L, pH, and the value is 2.5.
5. The method for purifying teicoplanin according to claim 1, wherein: in the step (1) and the step (2), the strong polar solvent is low-polyhydric alcohol; the buffer salt in the buffer salt water solution is one of sodium dihydrogen phosphate or potassium dihydrogen phosphate.
6. The method of claim 5, wherein the step of purifying teicoplanin comprises: the strong polar solvent is one of methanol or ethanol.
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