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A kind of rapid purification method of marine sulfated polysaccharide

The technology of a sulfated polysaccharide and a purification method is applied in the purification field of marine sulfated polysaccharide, which can solve the problems of inappropriate use in large quantities, low purification rate, and high loss of polysaccharide, and achieves the effects of saving purification time, good reproducibility, and simplified operation.

Active Publication Date: 2018-02-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Sevag method is most commonly used in the laboratory, but its disadvantages are: the process is long; the loss of polysaccharides is large; only a small amount of protein can be removed at a time, and it generally needs to be repeated many times to remove the protein; the solvent uses volatile chloroform, Large consumption, harmful to human body, pollutes the environment
Trifluorotrichloroethane method has higher deproteinization efficiency, but it is not suitable to be used in large quantities because of its volatility
However, the chromatographic column method is time-consuming and has a low purification rate (<25%), which cannot meet the needs of mass production

Method used

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  • A kind of rapid purification method of marine sulfated polysaccharide
  • A kind of rapid purification method of marine sulfated polysaccharide

Examples

Experimental program
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Embodiment 1

[0018] Embodiment 1: Rapid purification of philippine japonicus chondroitin sulfate:

[0019] 2g of philippine sea cucumber chondroitin sulfate crude polysaccharide through enzymolysis and 40% ethanol precipitation is completely dissolved in 50ml of phosphate buffered saline (pH6.3) to obtain an aqueous solution of sea cucumber sulfate crude polysaccharide; 200ml of QFF filler is filled in a 500ml sand core funnel ; Pour 50ml of crude polysaccharide solution into the sand core funnel, after the polysaccharide is adsorbed on QFF, elute 400ml with phosphate buffered saline, and connect to a vacuum pump for suction filtration; the mixture in the sand core funnel is then added to the sand core funnel with 0.1M Sodium chloride solution, elute 400ml, connect to the vacuum pump for suction filtration; add 1.5M sodium chloride solution to the mixture in the sand core funnel after suction filtration, connect to the vacuum pump for suction filtration, and elute 600ml in total; collect th...

Embodiment 2

[0020] Example 2: Rapid purification of fucoidan sulfate from Aposthopus japonicus:

[0021] 2g of philippine japonicus fucoidan sulfate crude polysaccharide precipitated by enzymolysis with 60% ethanol is completely dissolved in 50ml of phosphate buffered saline (pH6.3) to obtain an aqueous solution of sea cucumber sulfate crude polysaccharide; 400ml of QFF filler is filled in 500ml of sand In the core funnel; 50ml crude polysaccharide solution is poured into the sand core funnel, after the polysaccharide is adsorbed on QFF, 400ml is eluted with phosphate buffered saline, connected to a vacuum pump for suction filtration; the mixture in the sand core funnel is then poured into the sand core funnel Add 0.5M sodium chloride solution, elute 400ml, and connect to vacuum pump for suction filtration; add 3.8M sodium chloride solution to the mixture in the sand core funnel after suction filtration, connect to vacuum pump for suction filtration, and elute 600ml in total; collect the o...

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Abstract

The invention discloses a rapid purification method of marine sulfated polysaccharides. According to the method, firstly, crude marine sulfated polysaccharides are dissolved in phosphoric acid buffer salt and then poured into ion exchange resin filler, after the crude marine sulfated polysaccharides are adsorbed onto the filler, the crude marine sulfated polysaccharides are eluted with the phosphoric acid buffer salt, and vacuum filtration is performed; a low-concentration sodium chloride solution is added, and elution and vacuum filtration are performed; a high-concentration sodium chloride solution is added, and elution and vacuum filtration are performed; finally, centrifugation and alcohol precipitation are performed, freeze drying is performed after a product is dialyzed for one day, and the marine sulfated polysaccharides are obtained. The rapid purification method has the technical characteristics that the purification steps are simplified, the purification time is saved, and the purification efficiency is improved; no organic extraction solvents are introduced, the cost is reduced, and no environment pollution is caused; the method is stable, and the reproducibility is good; the equipment is simple, and the conditions are easy to control.

Description

technical field [0001] The invention relates to the technical field of purification of marine sulfated polysaccharides, in particular to a method for rapidly purifying sulfated polysaccharides by using QFF (QSepharose Fast Flow anion-exchange, agarose anion exchange resin) strong anion packing and a vacuum suction filtration device. Background technique [0002] Marine sulfated molluscs are rich in sulfated polysaccharides, and their structures often have highly repetitive structural sequences, which are easy to explore the structure-activity relationship and quality control. Among them, sea cucumber is one of the main sources of marine sulfated polysaccharides. There are mainly two types of sulfated polysaccharides in sea cucumbers: sea cucumber chondroitin sulfate (Sea Cucumber Chondroitin Sufate, SC-CHS) and sea cucumber fucoidan sulfate (sea cucumber Fucan, SC-FUC). Among them, sea cucumber chondroitin sulfate is an acidic mucopolysaccharide with fucose branched chains,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/08
CPCC08B37/0003C08B37/0069
Inventor 陈士国李珊李俊慧支梓鉴叶兴乾胡亚芹
Owner ZHEJIANG UNIV