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Method for efficiently separating immunocompetence polysaccharide of ganoderma atrum

An immune activity and polysaccharide technology, applied in the field of extraction and separation of natural products, can solve the problems of incomplete separation, time-consuming, poor stability, etc., and achieve the effect of shortening purification time, improving purification efficiency, and excellent immune activity

Inactive Publication Date: 2016-02-03
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Common gradient elution anion exchange chromatography can preliminarily separate polysaccharides in a relatively short period of time, but it is difficult to prepare polysaccharide components with uniform properties; and gel column chromatography is restricted due to long time-consuming and poor stability. Separation efficiency of polysaccharides, unable to prepare large quantities of bioactive polysaccharide fractions
The present invention uses Q-SepharoseFastFlow as the separation medium, and uses different concentrations of salt solutions as the eluent to carry out segmental elution, and attempts to provide a method for efficiently separating and preparing immune-active polysaccharides from black ganoderma lucidum, which can improve the purity of polysaccharides and enrich immune The role of active polysaccharide components, simple operation, good reproducibility, high efficiency, overcomes the shortcomings of gel column and other separation techniques such as low efficiency, long time-consuming, incomplete separation, etc.

Method used

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  • Method for efficiently separating immunocompetence polysaccharide of ganoderma atrum
  • Method for efficiently separating immunocompetence polysaccharide of ganoderma atrum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Weigh 60 mg of refined polysaccharide PSG sample, dissolve it in distilled water, and prepare a solution with a concentration of 3.0 mg / mL. Before sample injection, the sample solution is centrifuged (5000 rpm for 10 min) to remove insoluble matter; Put it into a Q-Sepharose FastFlow anion exchange column (1.5x10cm), let it seep down naturally, and fully exchange the sample with the resin for 10 minutes; Elute the unadsorbed samples with ultrapure water, and collect the eluent; the samples adsorbed on the resin are then eluted in stages with 0.1, 0.2, 0.5 and 2M NaCl solutions of 5 times the column volume, and the flow rate is controlled at 10mL The fractions of different stages were collected respectively; the eluents of five different fractions were concentrated, desalted by water dialysis, and freeze-dried to obtain separated and purified samples, which were recorded as Fw, Fw, and Fw respectively. 0.1 ,F 0.2 ,F 0.5 and F 2 , the sample recoveries were: 7.9%, 22.5...

Embodiment 2

[0026] Weigh 100 mg of the refined polysaccharide PSG sample, dissolve it in distilled water, and prepare a solution with a concentration of 5.0 mg / mL. Before sample injection, the sample solution is centrifuged (5000 rpm for 10 min) to remove insoluble matter; Put it into a Q-Sepharose FastFlow anion exchange column (1.5x10cm), let it seep down naturally, and fully exchange the sample with the resin for 10 minutes; Elute the unadsorbed samples with ultrapure water, and collect the eluent; the samples adsorbed on the resin are then eluted in stages with 0.1, 0.2, 0.5 and 2M NaCl solutions of 5 times the column volume, and the flow rate is controlled at 10mL The fractions of different stages were collected respectively; the eluents of five different fractions were concentrated, desalted by water dialysis, and freeze-dried to obtain separated and purified samples, which were recorded as Fw, Fw, and Fw respectively. 0.1 ,F 0.2 ,F 0.5 and F 2 , the sample recoveries were: 8.4%,...

Embodiment 3

[0028]Weigh 150 mg of refined polysaccharide PSG sample, dissolve it in distilled water, and prepare a solution with a concentration of 5.0 mg / mL. Before sample injection, the sample solution is centrifuged (5000 rpm for 10 min) to remove insoluble matter; Put it into a Q-Sepharose FastFlow anion exchange column (1.5x10cm), let it seep down naturally, and fully exchange the sample with the resin for 10 minutes; Elute the unadsorbed samples with ultrapure water, and collect the eluent; the samples adsorbed on the resin are then eluted in stages with 0.1, 0.2, 0.5 and 2M NaCl solutions of 5 times the column volume, and the flow rate is controlled at 10mL The fractions of different stages were collected respectively; the eluents of five different fractions were concentrated, desalted by water dialysis, and freeze-dried to obtain separated and purified samples, which were recorded as Fw, Fw, and Fw respectively. 0.1 , F 0.2 , F 0.5 and F 2 , the sample recoveries were: 8.1%, 22...

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Abstract

The invention discloses a method for efficiently separating immunocompetence polysaccharide of ganoderma atrum. Refined ganoderma atrum polysaccharide PSG serves as raw materials of the method, strong anion exchange resin Q-Sepharose Fast Flow is adopted for separating and purifying the PSG, sodium chloride solutions in different concentrations (0-2 M) are used for performing stepwise elution, different obtained components are sequentially dialyzed, frozen and dried, and ganoderma atrume acidic beta-(1->3, 1->6)-glucan polysaccharide components with immunocompetence are obtained. The method has the advantages that different chemical components in the PSG can be effectively separated by the strong anion exchange resin; the prepared acidic polysaccharide is high in sugar content, good in uniformity and superior to refined polysaccharide PSG in immunocompetence; the method is easy to operate, fast, efficient and good in reproducibility, elution liquid in use is free of toxin and pollution, and the method can be widely used for large-scale preparation of immunocompetence polysaccharide of ganoderma atrum.

Description

technical field [0001] The invention relates to the technical field of extraction and separation of natural products, in particular to a method for efficiently separating immune active polysaccharides from black ganoderma lucidum. Background technique [0002] Ganoderma lucidum is a precious medicinal fungus, which has the effects of nourishing and strengthening the body, and strengthening the body. Black Ganoderma lucidum is an important species in the genus Ganoderma lucidum. A large number of studies have shown that polysaccharides are one of the main functional components of Ganoderma lucidum. Black Ganoderma lucidum polysaccharides have been proven to have good biological activities such as immune regulation, anti-tumor, anti-oxidation, heart protection and anti-diabetes. Therefore, they are considered in the fields of medicine and health products It has broad development prospects. [0003] There are many methods for the separation and purification of polysaccharides,...

Claims

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Application Information

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IPC IPC(8): C08B37/00
Inventor 聂少平张汇谢明勇殷军艺
Owner NANCHANG UNIV
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