Application of monodispersed polymethacrylate ion exchange chromatography medium in column chromatography purification of fondaparinux sodium

A technology of polymethacrylate and ion-exchange chromatography, which is applied in organic chemistry, chemical instruments and methods, preparation of sugar derivatives, etc., can solve problems such as limited application and deformation, and achieve high purity, recovery, and high flow rate , the effect of high pressure flow rate

Active Publication Date: 2012-09-12
SUZHOU NANOMICRO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the medium used in this method is an agarose matrix, and its column volume will change under different flow rates, and compared with polyacrylate matrix, it will deform under higher pressure conditions, which limits its application in high flow rate separations. application

Method used

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  • Application of monodispersed polymethacrylate ion exchange chromatography medium in column chromatography purification of fondaparinux sodium
  • Application of monodispersed polymethacrylate ion exchange chromatography medium in column chromatography purification of fondaparinux sodium
  • Application of monodispersed polymethacrylate ion exchange chromatography medium in column chromatography purification of fondaparinux sodium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Purification of Fondaparinux Sodium:

[0022] A 26×310mm glass chromatographic column was used, and Uni Q-50S (produced by Suzhou Nan Microbe Technology Co., Ltd.) was used as the chromatographic packing material to fill the chromatographic column. The column volume was 164.5mL. The column was then equilibrated with 2.0 CV of ultrapure water at a flow rate of 10 mL / min. The crude product of fondaparinux sodium (purity 70%) was dissolved in ultrapure water, the concentration of the crude product was 4 mg / mL, and the sample volume was 250 mL. After sample loading, wash 2.0BV with ultrapure water first, then elute 3.5BV with 0.3M NaCl to remove impurities, then elute with 0.44M NaCl and collect the eluate in separate tubes, and finally elute with 1.0M NaCl until there is no UV absorption . After testing the collected liquid, the recovery rate of chromatographic purity above 99% is about 30%, and the recovery rate of chromatographic purity above 97% is about 60%.

Embodiment 2

[0024] Purification of Fondaparinux Sodium:

[0025] A 26×310mm glass chromatographic column was used, and Q Sepharose Fast Flow (GE Healthcare, particle size range 45-165 microns) was used to fill the chromatographic column with a column volume of 164.5mL. Activate the chromatographic column with NaCl, and then equilibrate the chromatographic column with 2.0CV ultrapure water at a flow rate of 10 mL / min. The crude product of fondaparinux sodium (purity 70%) was dissolved in ultrapure water, the concentration of the crude product was 4 mg / mL, and the sample volume was 250 mL. After loading the sample, wash 2.0BV with ultrapure water first, then elute 5BV with 0.36M NaCl to remove impurities, then elute with 0.6M NaCl and collect the eluate in separate tubes, and finally elute with 1.0M NaCl until there is no UV absorption. After testing the collected solution, the recovery rate of chromatographic purity above 99% is about 28%, and the recovery rate of chromatographic purity a...

Embodiment 3

[0027] Filling pressure performance test:

[0028] A 10×220mm glass chromatographic column was used, and Uni Q-50S (Suzhou Nan Microbe Technology Co., Ltd.) was used as the chromatographic filler. After the column was packed, the chromatographic column was equilibrated with 3CV ultrapure water at a flow rate of 1mL / min and the back pressure of the chromatographic column was measured. Start at this flow rate, gradually increase the flow rate, measure the back pressure of the chromatographic column every time the flow rate increases by 1mL / min, and make a flow rate-pressure curve, such as image 3 shown.

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Abstract

The invention discloses an application of a monodispersed polymethacrylate ion exchange chromatography medium in the column chromatography purification of fondaparinux sodium, wherein the column chromatography is carried out through adopting the monodispersed polymethacrylate ion exchange chromatography medium as the filler of a chromatographic column. The application is in favor of the separation and the purification of crude fondaparinux sodium with an aqueous solution as a mobile phase, and the reaching of high purity and high recovery rate, the column chromatography does not change with the change of the salt concentration and the salt flow velocity, the chromatography filler can resist high pressures and high flow velocities, and simultaneously the fondaparinux sodium adsorbed in the chromatography column can be eluted through the salt with a low concentration.

Description

technical field [0001] The invention relates to the field of medicine purification, in particular to the field of column chromatography purification of fondaparinux sodium. Background technique [0002] Fondaparinux sodium (Fondaparinux sodium) is the pentasaccharide substance of the smallest repeating segment of pentosan sodium. It is a specific inhibitor of activated coagulation factor Xa. It is mainly used for the prevention and treatment of thromboembolic diseases. It is widely used in To a certain extent, it solves the dilemma that anticoagulant drugs have plagued doctors all the time - finding a balance between anticoagulant intensity and bleeding risk. [0003] Fondaparinux sodium is artificially synthesized, and the general synthesis method needs to go through more than 50 steps, so the final crude product contains complex impurities, and the effective content of fondaparinux sodium is low (generally 60-80%). It is difficult to extract to high purity (above 98%). ...

Claims

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Application Information

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IPC IPC(8): C07H15/04C07H1/06
Inventor 江必旺陈荣姬吴俊成林生跃肖书霞
Owner SUZHOU NANOMICRO TECH CO LTD
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