CN115010792A - Purification method of vancomycin hydrochloride - Google Patents
Purification method of vancomycin hydrochloride Download PDFInfo
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- CN115010792A CN115010792A CN202210718612.9A CN202210718612A CN115010792A CN 115010792 A CN115010792 A CN 115010792A CN 202210718612 A CN202210718612 A CN 202210718612A CN 115010792 A CN115010792 A CN 115010792A
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- vancomycin
- purification method
- hydrochloric acid
- aqueous solution
- alumina
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- 108010059993 Vancomycin Proteins 0.000 title claims abstract description 56
- 229960001572 vancomycin hydrochloride Drugs 0.000 title claims abstract description 27
- LCTORFDMHNKUSG-XTTLPDOESA-N vancomycin monohydrochloride Chemical compound Cl.O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 LCTORFDMHNKUSG-XTTLPDOESA-N 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000000746 purification Methods 0.000 title claims abstract description 18
- 239000011347 resin Substances 0.000 claims abstract description 20
- 229920005989 resin Polymers 0.000 claims abstract description 20
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 44
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 claims description 30
- 229960003165 vancomycin Drugs 0.000 claims description 29
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 claims description 29
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 23
- 239000000243 solution Substances 0.000 claims description 23
- 239000007864 aqueous solution Substances 0.000 claims description 17
- 239000003480 eluent Substances 0.000 claims description 17
- 238000001914 filtration Methods 0.000 claims description 12
- 238000002425 crystallisation Methods 0.000 claims description 11
- 230000008025 crystallization Effects 0.000 claims description 11
- 238000004440 column chromatography Methods 0.000 claims description 9
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 8
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 5
- 238000012856 packing Methods 0.000 claims description 5
- 239000003963 antioxidant agent Substances 0.000 claims description 4
- 230000003078 antioxidant effect Effects 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000012043 crude product Substances 0.000 claims description 3
- 238000003795 desorption Methods 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 6
- 239000012467 final product Substances 0.000 abstract description 3
- 238000004587 chromatography analysis Methods 0.000 abstract description 2
- 239000002904 solvent Substances 0.000 abstract description 2
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 238000001035 drying Methods 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 7
- 238000001179 sorption measurement Methods 0.000 description 6
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 4
- MYPYJXKWCTUITO-KIIOPKALSA-N chembl3301825 Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)C(O)[C@H](C)O1 MYPYJXKWCTUITO-KIIOPKALSA-N 0.000 description 4
- 238000001728 nano-filtration Methods 0.000 description 4
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 229960003085 meticillin Drugs 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000006641 stabilisation Effects 0.000 description 3
- 238000011105 stabilization Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000191963 Staphylococcus epidermidis Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 description 1
- 229940001584 sodium metabisulfite Drugs 0.000 description 1
- 235000010262 sodium metabisulphite Nutrition 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K9/00—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
- C07K9/006—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
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a method for purifying vancomycin hydrochloride, which utilizes aluminum oxide and resin for two times of chromatography in sequence, has simple and convenient purification method, simple flow and less used solvent, and improves the purity and the stability of the product. The final product obtained by the purification method has the purity of over 98 percent and high stability, and can be stored for 24 months at normal temperature.
Description
Technical Field
The invention belongs to the technical field of medical products, and particularly relates to a purification method of vancomycin hydrochloride.
Background
Vancomycin hydrochloride is a glycopeptide antibiotic and is approved by FDA to be on the market in 1958. The medicament has the function characteristic of stronger bactericidal action on gram-positive bacteria, is clinically suitable for treating infection caused by methicillin-resistant staphylococcus aureus (MRSA) and other bacteria, and is a first choice medicament for treating diseases caused by methicillin-resistant staphylococcus aureus (MRSA) and methicillin-resistant staphylococcus epidermidis (MRSE).
The vancomycin hydrochloride production is firstly carried out with biological fermentation, and the separation and purification process comprises the following steps: pretreatment of fermentation liquor, ceramic membrane filtration, decolorization, chromatography, acidification crystallization, drying and other processes. Vancomycin hydrochloride produced by the existing process is mostly low in purity and poor in stability, and needs low-temperature storage and cold-chain transportation.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides a method for purifying vancomycin hydrochloride, which can improve the purity and stability of vancomycin hydrochloride and ensure that the product is suitable for normal-temperature storage and transportation.
The purification method of vancomycin hydrochloride provided by the invention comprises the following steps:
s1: preparing a vancomycin crude product into an aqueous solution, and adding hydrochloric acid to adjust the pH value to acidity to obtain a vancomycin hydrochloric acid aqueous solution A;
s2: carrying out alumina column chromatography on the vancomycin hydrochloric acid aqueous solution A obtained in the step S1, and collecting eluent;
s3: adsorbing the eluent obtained in the step S2 by using a resin column, resolving, collecting effluent, concentrating and crystallizing to obtain vancomycin;
s4: and (3) preparing the vancomycin crude product into an aqueous solution by using the vancomycin in the S3, adding hydrochloric acid to adjust the pH value to acidity to obtain a vancomycin hydrochloric acid aqueous solution B, filtering to obtain a filtrate, and crystallizing to obtain the vancomycin hydrochloride.
In the invention, because the existence of impurities leads the product to be easy to degrade and unstable, the invention aims at optimizing the internal quality of the product, improving the purity of the product and reducing the impurities.
In some embodiments of the present invention, the purity of the crude vancomycin at S1 is 85% or more.
In some embodiments of the invention, the concentration of the vancomycin hydrochloric acid aqueous solution A in S1 is 14g/L to 30 g/L.
Further, the concentration of the vancomycin hydrochloric acid aqueous solution A in the S1 is 15 g/L-27 g/L.
Furthermore, the concentration of the vancomycin hydrochloric acid aqueous solution A in the S1 is 21 g/L-24 g/L.
In some embodiments of the present invention, the pH of S1 is 2.5-4.5.
Further, the pH value of S1 is 3-4.
Further, the pH value of S1 is 3.3-3.7.
In some embodiments of the present invention, the alumina filler in the alumina column chromatography of S2 is selected from any one of neutral, weakly alkaline and weakly acidic alumina.
In some embodiments of the present invention, the alumina filler in the alumina column chromatography of S2 has a particle size of 80 mesh to 160 mesh.
In some embodiments of the invention, 1L of alumina filler is used per 20g to 25g of the crude vancomycin.
In some embodiments of the invention, the alumina column of S2 has a diameter to height ratio of 1: (3-6).
In some embodiments of the invention, the eluent for the alumina column chromatography described in S2 is water.
In some embodiments of the invention, the flow rate of the alumina column chromatography of S2 is 0.2BV/h to 0.5 BV/h.
Further, the flow rate is 0.3 BV/h-0.4 BV/h.
In some embodiments of the invention, the eluate of S2 is collected at a concentration > 600 mg/L.
In some embodiments of the invention, the resin packing in the resin column of S3 is a macroporous non-polar resin.
In some embodiments of the invention, the resin column of S3 has a diameter to height ratio of 1: (3-6).
In some embodiments of the invention, the volume of the resin packing in the resin column of S3 is 1/10 the volume of the alumina packing in the alumina column chromatography of S2.
In some embodiments of the present invention, the desorption solution desorbed from the resin column of S3 is water.
In some embodiments of the invention, the resin column of S3 adsorbs at a flow rate of 5BV/h to 6 BV/h.
In some embodiments of the invention, the resin column of S3 is resolved at a flow rate of 5BV/h to 6 BV/h.
In some embodiments of the invention, the effluent is collected at a concentration of > 400mg/L in S3.
In some embodiments of the invention, the concentration of S3 is concentrated using nanofiltration.
In some embodiments of the invention, the concentration of the concentrated solution obtained by the concentration of S3 is 25g/L to 55 g/L.
Further, the concentration of the concentrated solution obtained by the concentration in S3 is 30g/L to 50 g/L.
Further, the concentration of the concentrated solution obtained by the concentration in S3 is 35g/L to 45 g/L.
In some preferred embodiments of the present invention, the crystallizing of S3 comprises: adding antioxidant into the concentrated solution, adding ammonia water to adjust pH to alkalinity, and crystallizing.
In some preferred embodiments of the present invention, the antioxidant is selected from at least one of sodium sulfite, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, or disodium ethylenediaminetetraacetate.
In some preferred embodiments of the present invention, the antioxidant is added in an amount of 7g to 9g per liter of the solution.
In some preferred embodiments of the present invention, the concentration of the aqueous ammonia is 8% to 10%.
In some preferred embodiments of the present invention, the pH is adjusted to a basic value of 7.2 to 8.6 by adding ammonia water.
Further, adding ammonia water to adjust the pH value to be 7.6-8.0.
In some preferred embodiments of the present invention, the crystallization temperature of S3 is 20 ℃ to 30 ℃.
In some preferred embodiments of the present invention, the crystallization time of S3 is 4 to 8 hours.
In some more preferred embodiments of the present invention, the concentration of the vancomycin hydrochloride aqueous solution B of S4 is 80g/L to 160 g/L.
Furthermore, the concentration of the vancomycin hydrochloric acid aqueous solution B is 90 g/L-140 g/L.
Furthermore, the concentration of the vancomycin hydrochloric acid aqueous solution B is 110 g/L-120 g/L.
In some more preferred embodiments of the present invention, the vancomycin hydrochloride aqueous solution B of S4 has a pH of 3 to 4.
In some more preferred embodiments of the present invention, the addition of sodium chloride in S4 is preferably a sodium chloride solution.
In some more preferred embodiments of the invention, the concentration of the sodium chloride solution is 25% to 30%.
In some more preferred embodiments of the invention, the volume of the sodium chloride solution is 30% of the volume of the vancomycin aqueous hydrochloric acid solution B.
In some more preferred embodiments of the present invention, the crystallization time of S4 is 5 days to 10 days.
Further, the crystallization time of S4 is 7 to 9 days.
In some more preferred embodiments of the present invention, the crystallization temperature of S4 is 18 ℃ to 30 ℃.
In some more preferred embodiments of the present invention, the crystallization of S4 further comprises filtration to obtain vancomycin hydrochloride wet product, ethanol washing, and drying.
The beneficial effects of the invention are as follows:
the purification method is simple and convenient, the flow is simple, the used solvent is less, the purity of the vancomycin hydrochloride is improved, the stability of the vancomycin hydrochloride is improved, the purity of the final product is over 98 percent, and the final product can be stored for 24 months at normal temperature.
Detailed Description
The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention. The crude vancomycin can be obtained from fermentation liquor by using simple primary extraction and crystallization technology commonly used in the industry.
Example 1
In this embodiment, a vancomycin hydrochloride is purified, and the specific process is as follows:
(1) 50g of crude vancomycin is added with 2L of water, hydrochloric acid is added to adjust the pH value to 3.4, and insoluble substances are filtered out after stabilization for 30 min.
(2) The filtered solution was adsorbed by a 2L alumina column packed in advance with a diameter to height ratio of 1: 3. After adsorption, the elution is carried out by using water, the flow rate is controlled to be 600 mL/h-800 mL/h, and the eluent with the concentration of more than 600mg/L is collected.
(3) The alumina chromatographic eluent is absorbed by XBJ-152 resin, the volume of the resin is 200mL, the diameter-height ratio is 1:3, the flow rate is controlled to be 1000 mL/h-1200 mL/h, and the effluent liquid with the concentration of more than 400mg/L is collected.
(4) After adsorption, eluting with water, controlling the flow rate at 1000 mL/h-1200 mL/h, collecting the eluent with the concentration of more than 400mg/L, and combining the eluent with the previously collected effluent.
(5) The combined solution was concentrated by nanofiltration to a concentration of 38.4g/L and a volume of 1.08L.
(6) And adding 7.5g of sodium bisulfite into the concentrated solution, adding 8% ammonia water, adjusting the pH value to 7.9, crystallizing, controlling the temperature to be between 20 and 30 ℃, filtering after 8 hours, and drying to obtain 39.76g of vancomycin.
(7) And adding 330mL of water into the vancomycin obtained in the last step, adding hydrochloric acid to adjust the pH value to 3.42, stirring for 30min, and filtering.
(8) 99mL of 26 percent sodium chloride solution is added into the filtrate, and the mixture is kept stand and crystallized for 9 days at the temperature of 18-30 ℃.
(9) Filtering, washing with 160mL of ethanol, and drying to obtain vancomycin hydrochloride with the purity of 98.1%.
Stability data are monitored as shown in table 1 below:
TABLE 1
As can be seen from the above, the vancomycin hydrochloride obtained in the examples is stored at normal temperature for 24 months, and the purity is 92.3%.
Example 2
In this embodiment, a vancomycin hydrochloride is purified, and the specific process is as follows:
(1) 50g of crude vancomycin is added with 2L of water, hydrochloric acid is added to adjust the pH value to 3.7, and insoluble substances are filtered out after stabilization for 30 min.
(2) The filtered solution was adsorbed by a 2L alumina column packed in advance with a ratio of 1:4 in diameter to height. After adsorption, the elution is carried out by using water, the flow rate is controlled to be 600 mL/h-800 mL/h, and the eluent with the concentration of more than 600mg/L is collected.
(3) The alumina chromatographic eluent is absorbed by XBJ-318 resin, the volume of the resin is 200mL, the diameter-height ratio is 1:4, the flow rate is controlled to be 1000 mL/h-1200 mL/h, and the effluent liquid with the concentration of more than 400mg/L is collected.
(4) After adsorption, eluting with water, controlling the flow rate at 1000 mL/h-1200 mL/h, collecting the eluent with the concentration of more than 400mg/L, and combining the eluent with the previously collected effluent.
(5) The combined solution was concentrated by nanofiltration to a concentration of 35.2g/L and a volume of 1.13L.
(6) Adding 8g of sodium bisulfite into the concentrated solution, adding 8% ammonia water, adjusting the pH value to 7.7, crystallizing, controlling the temperature at 20-30 ℃, filtering after 8h, and drying to obtain 37.32g of vancomycin.
(7) Adding 310mL of water into the vancomycin obtained in the last step, adding hydrochloric acid to adjust the pH value to 3.67, stirring for 30min, and filtering.
(8) 93mL of 28 percent sodium chloride solution is added into the filtrate, and the mixture is kept stand and crystallized for 8 days at the temperature of 18-30 ℃.
(9) Filtering, washing with 180mL of ethanol, and drying to obtain vancomycin hydrochloride with the purity of 98.6%.
Stability data are monitored as shown in table 2 below:
TABLE 2
As can be seen from the above, the vancomycin hydrochloride of the embodiment is stored at normal temperature for 24 months, and the purity is 93.0%.
Example 3
In this embodiment, a vancomycin hydrochloride is purified, and the specific process is as follows:
(1) 50g of crude vancomycin is added with 2L of water, hydrochloric acid is added to adjust the pH value to 3.3, and insoluble substances are filtered after stabilization for 30 min.
(2) The filtered solution was adsorbed by a 2L alumina column packed in advance with a ratio of 1: 5. After adsorption, the elution is carried out by using water, the flow rate is controlled to be 600 mL/h-800 mL/h, and the eluent with the concentration of more than 600mg/L is collected.
(3) The alumina chromatographic eluent is absorbed by XBJ-318 resin, the volume of the resin is 200mL, the diameter-height ratio is 1:4, the flow rate is controlled between 1000mL/h and 1200mL/h, and the effluent liquid with the concentration of more than 400mg/L is collected.
(4) After adsorption, eluting with water, controlling the flow rate at 1000 mL/h-1200 mL/h, collecting the eluent with the concentration of more than 400mg/L, and combining the eluent with the previously collected effluent.
(5) The combined solution was concentrated by nanofiltration to a concentration of 34.1g/L in a volume of 1.25L.
(6) Adding 8.5g of sodium bisulfite into the concentrated solution, adding 10% ammonia water, adjusting the pH value to 8.0, crystallizing, controlling the temperature at 20-30 ℃, filtering after 8h, and drying to obtain 40.37g of vancomycin.
(7) Adding 340mL of water into the vancomycin obtained in the last step, adding hydrochloric acid to adjust the pH value to 3.35, stirring for 30min, and filtering.
(8) 102mL of 26% sodium chloride solution is added into the filtrate, and the mixture is kept stand and crystallized for 7 days at the temperature of 18-30 ℃.
(9) Filtering, washing with 200mL of ethanol, and drying to obtain vancomycin hydrochloride with the purity of 98.3%.
Stability data are monitored as follows:
TABLE 3
As can be seen from the above, the vancomycin hydrochloride of the embodiment is stored at normal temperature for 24 months, and the purity is 93.0%.
The embodiments of the present invention have been described in detail, but the present invention is not limited to the embodiments, and various changes can be made without departing from the gist of the present invention within the knowledge of those skilled in the art. Furthermore, the embodiments of the present invention and features of the embodiments may be combined with each other without conflict.
Claims (10)
1. A purification method of vancomycin hydrochloride is characterized by comprising the following steps:
s1: preparing a vancomycin crude product into an aqueous solution, and adding hydrochloric acid to adjust the pH value to acidity to obtain a vancomycin hydrochloric acid aqueous solution A;
s2: carrying out alumina column chromatography on the vancomycin hydrochloric acid aqueous solution A in S1, and collecting eluent;
s3: adsorbing the eluent in the S2 by using a resin column, resolving, collecting effluent liquid, concentrating and crystallizing to obtain vancomycin;
s4: preparing vancomycin in the S3 into an aqueous solution, adding hydrochloric acid to adjust the pH value to acidity to obtain a vancomycin hydrochloric acid aqueous solution B, filtering to obtain a filtrate, and crystallizing to obtain vancomycin hydrochloride.
2. The purification method according to claim 1, wherein the pH value in S1 is 2.5 to 4.5.
3. The purification method of claim 1, wherein the purity of the crude vancomycin in S1 is not less than 85%.
4. The purification method according to claim 1, wherein the volume mass ratio of the alumina filler to the crude vancomycin in the alumina column chromatography is 1L: (20-25) g.
5. The purification method according to claim 4, wherein the flow rate of the alumina column chromatography in S2 is 0.2BV/h to 0.5 BV/h.
6. The purification process of claim 4, wherein the volume of resin packing in the resin column in S3 is 1/10 of the volume of the alumina packing.
7. The purification method according to claim 1, wherein the flow rate of the desorption in S3 is 5BV/h to 6 BV/h.
8. The purification method according to claim 1, wherein the crystallization in S3 comprises the steps of adding an antioxidant to the concentrated solution, adjusting the pH to alkaline with ammonia water, and crystallizing.
9. The purification process according to claim 1, wherein the crystallization time in S3 is 4 to 8 hours.
10. The purification process according to claim 1, wherein the crystallization time in S4 is 5 to 10 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210718612.9A CN115010792A (en) | 2022-06-23 | 2022-06-23 | Purification method of vancomycin hydrochloride |
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| US20060003406A1 (en) * | 2004-06-30 | 2006-01-05 | Lee Ju W | Process of purifying vancomycin hydrochloride |
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