Method for radial flow chromatogram to remove protein in crude polysaccharide

A crude polysaccharide and protein technology, applied in the field of removing protein from crude polysaccharides, can solve the problems of large consumption of organic solvents, injury to operators, long operation cycle, etc., and achieve convenient follow-up processing, less environmental pollution, and short operation time Effect

Inactive Publication Date: 2009-06-03
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Existing methods for removing protein from crude polysaccharides have problems such as huge consumption of organic solvents, serious loss of polysaccharides, high cost, long operating cycle, envir

Method used

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  • Method for radial flow chromatogram to remove protein in crude polysaccharide

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Experimental program
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Effect test

Embodiment 1

[0026] Take the dried and crushed rapeseed meal, add water according to the ratio of rapeseed meal to water mass ratio of 1:20, extract at 95℃ for 2 hours, filter, concentrate the filtrate, precipitate the concentrate with alcohol, centrifuge, and take the precipitate Rapeseed crude polysaccharide is obtained by vacuum freeze-drying.

Embodiment 2

[0028] Take the dried and crushed rapeseed meal, add water according to the ratio of rapeseed meal to water mass ratio of 1:30, extract at 100°C for 4 hours, filter, concentrate the filtrate, precipitate the concentrate with alcohol, centrifuge, and take the precipitate Rapeseed crude polysaccharide is obtained by vacuum freeze-drying.

Embodiment 3

[0030] (1) Instruments and reagents:

[0031] Radial flow chromatography column (volume 250ml, Sepragen, USA), constant flow pump (BT00-300M, Baoding Lange Constant Flow Pump Co., Ltd.), beaker;

[0032] C104E, C107E weakly acidic cation exchange resins (provided by Deqing Bolaite (China) Co., Ltd.);

[0033] Rapeseed crude polysaccharide (obtained in Examples 1 and 2), Ganoderma lucidum polysaccharide (provided by Zhejiang Yisheng Fungus Development Co., Ltd.);

[0034] Hydrochloric acid solution, sulfuric acid solution, sodium hydroxide and potassium hydroxide are all analytically pure.

[0035] (2) The specific operation process is as follows:

[0036] 1. Resin pretreatment: Soak the C104E cation exchange resin provided by Deqing Bleachlight (China) Co., Ltd. in a 3% hydrochloric acid solution for 4 hours, wash with pure water to neutrality, and then place the resin in a 2% NaOH solution Soak for 2 hours, wash with pure water to neutral, and finally soak the resin in 3% hydroch...

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Abstract

The invention discloses a method for radial flow chromatogram to remove protein in crude polysaccharide. In the method, the crude polysaccharide containing the protein is confected as aqueous solution of 10-30mg/ml to be centrifugated to get supernatant fluid; the supernatant fluid passes through a radial flow chromatogram column which is filled with cation resin and is eluted by pure water as eluent; sample-loading flow speed is controlled at 1-3ml/min; eluting flow speed is 5-20ml/min; elution solution is collected until no sugar is detected in the elution solution; and the collected elution solution is combined to be carried out post treatment to obtain pure polysaccharide. The method has short operation time, large sample processing capacity, high protein-removing rate, less polysaccharide loss and wide application range and can be used for removing the protein from plant polysaccharide and fungal polysaccharide.

Description

(1) Technical field [0001] The invention belongs to the technical field of polysaccharide separation and purification, and specifically relates to a method for removing proteins in crude polysaccharides. (2) Background technology [0002] As a biologically active product, polysaccharides generally need to be separated and purified to a certain purity before they can play a better role, especially when it is to be developed into drugs with higher purity requirements, it is essential to remove the impurity protein contained in it.的purification step. For polysaccharide injection products, it is necessary to remove the protein in the polysaccharide, otherwise the protein will easily become a heat-sensitive source. In addition, for further analysis and identification of the polysaccharide structure, it is also required that the protein must be removed. Active polysaccharides are mainly natural plant polysaccharides and some fungal polysaccharides. These polysaccharide components have ...

Claims

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Application Information

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IPC IPC(8): C08B37/00
Inventor 孙培龙邵平何晋浙姜慧燕
Owner ZHEJIANG UNIV OF TECH
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