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Pseudomonas elodea rhzomorph purifying method

The technology of a pseudomonosporin and a purification method is applied in the field of purification of pseudomonosporins, which can solve the problems of long time and low purity, and achieve the effects of easy control and simplified process technology.

Inactive Publication Date: 2015-02-25
江苏汉邦科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a new method for purifying high-purity pseudomycin by using a liquid chromatography system for the long time and low purity problems existing in the process of purifying pseudomycin. The method includes the following steps:

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] 1. Take 1 g of the pseudomycin fermentation broth and dissolve the crude product in water to prepare a saturated solution, place it in a filter device, and filter to remove solid particles;

[0010] 2. Pump the pseudomycin solution into the dynamic axial compression column to prepare the chromatographic system. The column packing size is Φ50×250mm, the filler is octadecyl silica gel, the particle size is 10~30um, and the sample volume is 1g. The volume ratio of the mobile phase is 30:70. After the pseudomycin is eluted, the mobile phase is switched to acetonitrile with a high proportion to wash out the rear-end impurities, and a separation cycle ends. The detection wavelength of the ultraviolet-visible photometric detector used is 240nm, and the fractions with a retention time of 35-49min are collected, and the purity is ≥98.8% by HPLC analysis.

Embodiment 2

[0012] 1. Take 1.5g of the pseudomycin fermented liquid and dissolve the crude product in water to prepare a saturated solution, place it in a filter device, and filter to remove solid particles;

[0013] 2. Pump the pseudomycin solution into the dynamic axial compression column to prepare the chromatographic system. The column packing size is Φ50×250mm, the filler is octadecyl silica gel, the particle size is 10~15um, and the sample volume is 1.5g. The volume ratio of the mobile phase is 26:74. After the elution of the pseudomycin is completed, the mobile phase is switched to acetonitrile with a high proportion to flush out the rear-end impurities, and a separation cycle ends. The detection wavelength of the ultraviolet-visible photometric detector used is 240nm, and the fractions with a retention time of 31-47min are collected, and the purity is ≥98.5% by HPLC analysis.

Embodiment 3

[0015] 1. Take 3g of the pseudomycin fermentation broth and dissolve the crude product in water to prepare a saturated solution, place it in a filter device, and filter to remove solid particles;

[0016] 2. Pump the pseudomycin solution into the dynamic axial compression column to prepare the chromatographic system. The column packing size is Φ150×250mm, the filler is octadecyl silica gel, the particle size is 10um, and the sample volume is 1g. The volume ratio of the mobile phase is 22:78. After the elution of pseudomycin is completed, the mobile phase is switched to acetonitrile with a high proportion to flush out the rear-end impurities, and a separation cycle ends. The detection wavelength of the ultraviolet-visible photometric detector used was 240nm, and the fractions with a retention time of 42-53min were collected, and the purity was ≥98.1% by HPLC analysis.

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Abstract

The invention discloses a pseudomonas elodea rhzomorph purifying method, pseudomonas elodea rhzomorph fermentation liquor is used as a raw material, appropriate column separation conditions can be obtained by optimization of conditions, and the appropriate column separation conditions are further enlarged onto a dynamic axial compression industrial preparative chromatography system for separation for obtaining of high purity pseudomonas elodea rhzomorph. The advantages of the method are that, good effect of separation and purification can be achieved by direct use of the dynamic axial compression industrial preparative chromatography for separation of an extracted crude product, the method has the advantages of simple operation, short period and high efficiency, and the purity can reach more than 98.5%.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for purifying pseudomycin. technical background [0002] Pseudomycin, drug name mupirocin, is an inhibitor of aminoacyl-tRNA synthetase (AaRS), suitable for skin and soft tissue infections caused by Gram-positive cocci, such as impetigo, boils, folliculitis Primary infection and secondary infection of eczema, dermatitis, ulcer, trauma and other skin diseases. Purification of pseudomycin products by traditional chromatography takes a long time and the purity is not high. The present invention uses high-pressure preparation chromatography to solve these problems and achieves a satisfactory separation effect. Therefore, this method can be used in the research of natural products and synthetic medicines. effective means. Contents of the invention [0003] The purpose of the present invention is to provide a new method for purifying high-purity pseudomycin...

Claims

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Application Information

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IPC IPC(8): C07D407/06
CPCC07D407/06
Inventor 张宇张大兵刘根水
Owner 江苏汉邦科技股份有限公司
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