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Method for purifying tiacumicin B by high performance liquid chromatography

A high-performance liquid chromatography and taigamycin technology, which is applied in the field of high-performance liquid chromatography to purify taigamycin B, can solve the problems that the separation effect needs to be further improved, the product purity cannot meet the requirements, the separation factor is low, and the like. High recovery rate, good removal effect, high purity effect

Active Publication Date: 2013-03-27
SUZHOU NANOMICRO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In these existing technologies, there are problems such as the low separation factor of the purification medium and poor column efficiency, so that the purity of the product cannot meet the requirements and the yield is low, and the separation effect needs to be further improved.

Method used

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  • Method for purifying tiacumicin B by high performance liquid chromatography
  • Method for purifying tiacumicin B by high performance liquid chromatography
  • Method for purifying tiacumicin B by high performance liquid chromatography

Examples

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Effect test

Embodiment 1

[0026] Purification of tiacumicin B: use a 15×310mm glass column, Uni30BPC resin (pyrrolidone and styrene copolymer particle size 30±1.5um, Suzhou Nano Microorganism Technology Co., Ltd.) as chromatography filler, and the column volume is 55ml , wash 2 CVs (column volumes) with 60% acetonitrile in water, and then equilibrate 2 CVs with 20% acetonitrile. Mobile phase: A: 0.1% formic acid aqueous solution; B: acetonitrile. The crude tiacumicin B (purity 78%) was dissolved in 60% methanol aqueous solution and filtered with a 0.45um filter membrane. The concentration is 8mg / ml, the sample volume is 45.8ml, the sample flow rate is 5ml / min, 1CV of 20% B is pre-washed, and then 8 CVs of 50% B are eluted. Collected in sections, the purity of the collected liquid in the front section is 85.6% (basically no after-impurity), accounting for 20.9% of the sample volume; the purity of the middle section is 90.72%, accounting for 49.85% of the sample volume; Miscellaneous), accounting for 2...

Embodiment 2

[0030] Purification of tiacumicin B: use 15×310mm glass column, Uni30BPC resin (material pyrrolidone and styrene copolymer ball particle size 30±1.5um, Suzhou Nano Microbe Technology Co., Ltd.) as chromatography packing material, the column volume is 55ml, wash 2 CVs with 60% acetonitrile aqueous solution first, then equilibrate 2 CVs with 20% acetonitrile. Mobile phase: A 0.1% formic acid in water; B acetonitrile. The collected front stage tiacumicin B was mixed in 5 batches and then concentrated under reduced pressure to make the concentration of acetonitrile lower than 35%. Samples were loaded directly, the flow rate was 5ml / min, 1CV of 20% B was pre-washed, and then 8 CVs of 45% B were eluted. Collected in sections, the yield with a purity of 98% was 89.82%.

Embodiment 3

[0032] Purification of tiacumicin B: use 15×310mm glass column, Uni30BPC resin (material pyrrolidone and styrene copolymer ball particle size 30±1.5um, Suzhou Nano Microbe Technology Co., Ltd.) as chromatography packing material, the column volume is 55ml, wash 2 CVs with 60% acetonitrile aqueous solution first, then equilibrate 2 CVs with 20% acetonitrile. Mobile phase: A 0.1% aqueous formic acid; B acetonitrile. The collected tiacumicin B in the middle stage was mixed and then concentrated under reduced pressure to keep the concentration of acetonitrile below 35%. Direct sample loading, flow rate 5ml / min, 1 CV of 20% B prewash, and then 8 CVs of 50% B elution. Collected in sections, the yield with a purity of 98% was 36.46%.

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Abstract

The invention provides a method for purifying tiacumicin B by high performance liquid chromatography, wherein in the method, Uni30BPC monodisperse polymer microspheres are used as fillers for performing column chromatography in a purification and chromatography column. The invention provides a method which is better in separation effect for tiacumicin B, wherein a novel medium which is high in separation factor and column efficiency is adopted, thus overcoming the shortages of low separation factor and bad column efficiency existing in the prior art, having a good removal effect for various impurities in tiacumicin B, and being high in preparation purity and recovery rate.

Description

technical field [0001] The invention relates to a purification method, in particular to a method for purifying tiacumicin B by high performance liquid chromatography. Background technique [0002] Tiacumicins is a general term for a series of 18-membered ring macrolide antibiotics produced by the actinomycete Dactylosporangium aurantiacumsubsp.hamdenensis NRRL 18085. Tiacumicin has activity against various Gram-positive bacteria, especially against Clostridium difficile and Mycobacterium tuberculosis, and it also has antitumor activity. The study found that tiacumicn B is the best active component in tiacumicin, which has two deoxy sugar groups and an aromatic ring branch, C-18 is R-hydroxyl, and has a lower minimum inhibitory concentration value for Clostridium difficile . Tiacumicin B refers to a molecule with the following structure: [0003] . [0004] Tiacumicin B is almost insoluble in water because of its strong hydrophobicity, so it is generally prepared by rev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/08C07H1/06
Inventor 江必旺陈荣姬潘毅吴俊成
Owner SUZHOU NANOMICRO TECH CO LTD
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