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Separation and purification method of vancocin

A technology of vancomycin and vancomycin hydrochloride, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as difficult filtration, and achieve the effect of simple method and high yield

Active Publication Date: 2009-06-17
ZHEJIANG MEDICINE CO LTD XINCHANG PHAMACEUTICAL FACTORY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Separation of vancomycin with isopropanol, ethanol, or acetone results in a slurry of vancomycin that is very difficult to filter

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of vancomycin hydrochloride aqueous solution

[0026] 200L of vancomycin fermentation broth containing 6.8mg / ml of vancomycin is used for 2m under alkaline conditions pH=8.5~10.5 2 The clarified filtrate is filtered with a ceramic membrane of 0.02~0.5μm, and the obtained clarified filtrate is passed through 40L macroporous adsorption resin AMBERLITE XAD-1600. Vancomycin is adsorbed on the resin. The resin is washed with 200L purified water and then washed with 10L containing 40% ethanol and 60% 1mol / L hydrochloric acid aqueous solution was eluted, vancomycin was eluted, and 8.5L of eluate containing vancomycin was obtained, and the vancomycin concentration was 18.2mg / ml. 85 g of activated carbon was added to the eluate for decolorization, and the filtrate obtained by filtration was concentrated to obtain 1.05 L of an aqueous solution of vancomycin hydrochloride, in which the vancomycin concentration was 118 mg / ml and the chromatographic purity of van...

Embodiment 2

[0027] Example 2: Preparation of crude vancomycin

[0028] Take the volume of 200 mL of vancomycin hydrochloride water in Example 1, containing 80.4% of vancomycin chromatographic purity (HPLC), pH of the solution is 3.0, add NH while stirring 4 HCO 3 A total of 16.0g, during the process of adding salt, the solution continuously generates bubbles, vancomycin is precipitated from it, and a slurry is formed, and then concentrated ammonia is added to adjust the pH to 8.0. At this time, more vancomycin is precipitated and the slurry is thicker. Continue stirring for 50 minutes to make it evenly mixed, stop stirring and let it stand for 18 hours, and the process temperature is controlled at 19-20°C. The mixture was separated by filtration and top-washed with 50 mL of ethanol to obtain 38.2 g of a wet product of vancomycin base with a titer of 564 ug / mg and a yield of 91.3%. The chromatographic purity of vancomycin containing vancomycin was 88.6% detected by HPLC.

Embodiment 3

[0029] Example 3: Preparation of crude vancomycin

[0030] Take the volume of 200 mL of vancomycin hydrochloride water in Example 1, containing 80.4% of vancomycin chromatographic purity (HPLC), pH of the solution is 3.0, add NH while stirring 4 HCO 3 A total of 16.0g, during the process of adding salt, the solution continuously produces bubbles, vancomycin is precipitated from it, forming a slurry, and then concentrated ammonia is added to adjust the pH to 7.5. At this time, more vancomycin is precipitated and the slurry is thicker. Continue stirring for 45 minutes to make it evenly mixed, stop stirring and let it stand for 18 hours, and the process temperature is controlled at 14.5~15.5°C. The mixture was separated by filtration and top-washed with 50 mL ethanol to obtain 43.3 g of a wet product of vancomycin base with a titer of 505 ug / mg and a yield of 92.7%. The chromatographic purity of vancomycin containing vancomycin was 86.2% detected by HPLC.

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PUM

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Abstract

The invention relates to a method for separating vancomycin. The method comprises the following steps: (1) carrying out the column chromatography elution on the vancomycin fermentation liquor in a hydrochloric acid aqueous solution mobile phase macropore absorptive resin chromatographic column containing alcohol so as to obtain vancomycin aqueous solution; (2) adding NH4HCO3 into the hydrochloricacid vancomycin aqueous solution to form precipitate, adjusting the pH value to between 7.5 and 8.5 through ammonia water, and stirring and standing the solution; and (3) obtaining a vancomycin crudeproduct with the content of vancomycin chromatographic purity not less than 80 percent through filtration separation and alcohol top washing. The method is simple and has high yield. The content of the vancomycin chromatographic purity is not less than 80 percent; and the titer of a vancomycin wet product at least reaches 500 mu g / mg.

Description

Technical field [0001] The invention relates to a method for separating vancomycin. Specifically, in this method, the vancomycin fermentation broth is eluted by column chromatography in a macroporous adsorption resin chromatography column containing an ethanol-containing hydrochloric acid aqueous solution mobile phase, and then decolorization and nanofiltration are performed to obtain an aqueous vancomycin hydrochloride solution. At a certain temperature, add NH to the vancomycin hydrochloride aqueous solution 4 HCO 3 A precipitate is formed and adjusted with ammonia water to obtain a crude vancomycin with a chromatographic purity of not less than 80%. Background technique [0002] Vancomycin is mainly used to treat severe infections caused by strains of Staphylococcus that are resistant to β-endolactamase. For susceptible patients who are allergic to penicillin, vancomycin is an alternative antibiotic. Vancomycin can also treat postoperative diarrhea caused by refractory intesti...

Claims

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Application Information

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IPC IPC(8): C07K9/00C07K1/36C07K1/22
Inventor 王小勇孙新强
Owner ZHEJIANG MEDICINE CO LTD XINCHANG PHAMACEUTICAL FACTORY
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