Method for separating and purifying reduced glutathione

A technology of oxidized glutathione and glutathione, which is applied in the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problems of small differences in polarity, poor stability, easy to exceed the standard and other problems, and achieve operation Simple, mild reaction conditions and low energy consumption

Active Publication Date: 2016-05-11
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, GSH has poor stability in aqueous solution, and it is easy to degrade and oxidize during the ion exchange process. Therefore, low temperature and nitrogen protection are required in a closed environment to keep GSH from degrading and oxidizing; Impuritie

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Take 1 L of the GSH conversion solution by the enzymatic method, and the GSH content is 13.1 g. Add 3 g of potassium bromate, stir at room temperature for 15 h, adjust the pH to 3.5 with sulfuric acid, and centrifuge. Put the strong acid cation exchange resin HD-8 on the column, wash 5 times the column volume with 0.03mol / L acetic acid, and then elute with 5% sulfuric acid, and collect the eluate with a GSSG concentration higher than 0.2g / L. Put the non-polar adsorption resin SP207 on the column, elute with deionized water, and collect the eluate whose GSSG concentration is greater than 0.2g / L, wherein the GSSG content is 11.84g. Concentrate to 500 mL by nanofiltration, add 0.06 mol of zinc powder to the concentrated solution, ventilate and stir at room temperature, add 120 mL of 1 mol / L sulfuric acid dropwise, react for 1 hour, and centrifuge the reaction solution. Put the centrifuged solution on the non-polar adsorption resin SP207 column, elute with deionized water,...

Embodiment 2

[0062] Take 2L of yeast fermentation extract, with a GSH content of 11.2g. Add 3 g of potassium bromate, stir at room temperature for 15 h, adjust the pH to 3.5 with sulfuric acid, and centrifuge. Put the strong acid cation exchange resin HD-8 on the column, wash 5 times the column volume with 0.03mol / L acetic acid, and then elute with 5% sulfuric acid, and collect the eluate with a GSSG concentration higher than 0.2g / L. Put the non-polar adsorption resin SP207 on the column, elute with deionized water, and collect the eluate whose GSSG concentration is greater than 0.2g / L, wherein the GSSG content is 9.74g. Concentrate to 500 mL by nanofiltration, add 0.05 mol of zinc powder to the concentrated solution, ventilate and stir at room temperature, add 100 mL of 1 mol / L sulfuric acid dropwise, react for 1 hour, and centrifuge the reaction solution. Put the centrifuged solution on the non-polar adsorption resin SP207 column, elute with deionized water, and collect the eluate whose...

Embodiment 3

[0064] Take 1 L of the GSH conversion solution obtained by the enzymatic method, with a GSH content of 12.1 g. Add 3 g of potassium chlorate, stir at room temperature for 15 h, adjust the pH to 3.5 with sulfuric acid, and centrifuge. Put the strong acid cation exchange resin HD-8 on the column, wash 5 times the column volume with 0.03mol / L acetic acid, and then elute with 5% sulfuric acid, and collect the eluate with a GSSG concentration higher than 0.2g / L. Put the non-polar adsorption resin SP207 on the column, elute with deionized water, and collect the eluate whose GSSG concentration is greater than 0.2g / L, wherein the GSSG content is 9.56g. Concentrate to 500 mL by nanofiltration, add 0.05 mol of zinc powder to the concentrated solution, ventilate and stir at room temperature, add 100 mL of 1 mol / L sulfuric acid dropwise, react for 1 hour, and centrifuge the reaction solution. Put the centrifuged solution on the non-polar adsorption resin SP207 column, elute with deionize...

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Abstract

The invention provides a method for separating and purifying reduced glutathione from a solution containing glutathione. The method includes the steps of firstly, adding an oxidizing agent to the glutathione solution so that glutathione can be oxidized into oxidized glutathione; secondly, adding acid to the solution, regulating pH to 1.0-4.0, and conducting centrifugation or filtration; thirdly, processing the centrifuged or filtered solution obtained in the second step through ion exchange resin and adsorption resin, and obtaining oxidized glutathione eluant; fourthly, reducing the eluant obtained in the third step through a reducing agent to obtain a glutathione solution, desalting the reduced glutathione solution through adsorption resin, and conducting concentration, crystallization and drying to obtain glutathione. The method is easy and convenient to operate, low in pollution, free of low temperature and nitrogen protection, low in energy consumption, mild in reaction condition and suitable for industrialized production, and the purity of the obtained solid reduced glutathione is larger than 99%.

Description

technical field [0001] The invention relates to the field of medicine separation and purification, in particular to a method for separation and purification of reduced glutathione. Background technique [0002] Glutathione (GSH) is an important drug that regulates physiological functions. It has been widely used clinically and has become one of the most important drugs in medicine that can regulate human immune function and assist anti-cancer functions. . The antioxidant properties of GSH make its application in the food industry attract people's attention. Glutathione is a biologically active tripeptide compound containing γ-glutamyl and thiol, which is condensed from glutamic acid, cysteine, and glycine. It is used in the synthesis of proteins and DNA, the transport of amino acids, and cell It plays a direct or indirect role in important biological functions such as the protection of human body. It is mainly distributed in animals, plants, and microbial cells, and is wi...

Claims

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Application Information

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IPC IPC(8): C07K5/037C07K1/18
CPCC07K5/0215
Inventor 杨峰晨田慧杨勇卢泓宇徐期马骏张永进王应朋
Owner ZHEJIANG HISUN PHARMA CO LTD
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