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Extraction method of natural soluble polysaccharide using radial chromatography

A chromatographic purification, soluble technology, applied in the biological field

Inactive Publication Date: 2011-11-23
TIANJIN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the mid-1980s, radial chromatography technology began to be industrialized, mainly used in the separation and purification of proteins, such as the purification of monoclonal antibodies, the separation and purification of IGase enzymes, and the separation and purification of human urinary kallikrein. There are few reports on the application of chromatography to the separation and purification of polysaccharides

Method used

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  • Extraction method of natural soluble polysaccharide using radial chromatography
  • Extraction method of natural soluble polysaccharide using radial chromatography
  • Extraction method of natural soluble polysaccharide using radial chromatography

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0025] (1). Use DEAE-650M anion exchange resin to fill the radial chromatographic column (see figure 1 ), weighed 15 mg of hair vegetable crude polysaccharide and dissolved it in deionized water to form a polysaccharide solution with a concentration of 0.5 mg / mL. After the precipitate was removed by centrifugation, ion exchange chromatography was carried out.

[0026] (2). Carry out linear gradient elution from 01.0mol / L with 1000mL NaCl solution, the elution rate is 1mL / min, and the eluate is collected by a partial collector.

[0027] (3). Use the phenol-sulfuric acid method to detect the absorbance A of each tube 490 , taking the elution volume as the abscissa, and the absorbance value and the concentration gradient change of the NaCl solution as the ordinate to make a graph, the ion exchange chromatography elution curve of the polysaccharide, see figure 2 . It can be seen from the figure that the elution curve presents three independent peaks, that is, the polysaccharide...

example 2

[0030] (1). Use gel G-100 to fill the self-made radial chromatography column (see figure 1 ), take 3mg of Nostocyst polysaccharide (the Nostocyst polysaccharide of the 1st component that obtains in embodiment 1), be dissolved in deionized water and be made into the polysaccharide solution that concentration is 0.5mg / mL, after centrifugation removes precipitation, carry out gelation Chromatography to remove most of the electrolyte NaCl contained.

[0031] (2). Use 200mL deionized water for elution, the elution rate is 0.3mL / min, and use a partial collector to collect the eluate.

[0032] (3). Use the phenol-sulfuric acid method to detect the absorbance A of each tube 490 , taking the elution volume as the abscissa, the absorbance value and the conductivity change of NaCl as the ordinate, the gel chromatography elution curve of the polysaccharide (see image 3 ).

[0033] (4). The eluents before the intersection of the two curves were combined, concentrated under reduced pres...

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Abstract

The invention relates to a method for purifying naturally soluble polysaccharides by radial chromatography. After rough polysaccharide solution enters into a radial chromatographic column, the rough polysaccharide solution flows to the center from the periphery of the chromatographic column by flowing through a stationary phase filled by the chromatographic column, corresponding solvent is taken as a mobile phase for elution, and then the solution flows out from a central liquid outlet. As the radial chromatographic column is characterized in that samples have a contact area with filler at the starting position and gather at the liquid outlet, the method can separate or purify the naturally soluble polysaccharides.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for radial chromatographic purification of natural soluble polysaccharides. Background technique [0002] As early as the 1950s, with the continuous research of chromatography technology, radial chromatography developed rapidly. This chromatographic technique is mainly evolved from the traditional axial chromatographic technique by Rhee et al. At present, this chromatographic technique has been widely used in molecular separation techniques. [0003] In the relevant reports comparing radial chromatography with axial chromatography, radial chromatography has shown the advantages of large processing capacity and fast speed, and also has the advantages of large contact area between the sample and the filler at the beginning and convergence at the outlet. features. In the mid-1980s, radial chromatography technology began to be industrialized, mainly used in the separation and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D15/08C08B37/00
Inventor 贾士儒戴玉杰王静文谭之磊岳思君姚瑾郭伟许鹏贾梦瑶
Owner TIANJIN UNIV OF SCI & TECH
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