Method for separating and purifying reduced glutathione (GSH) from reduced glutathione contained fermentation leaching liquid

A technology for separation and purification of glutathione, which is applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as no mention of purity information, and achieve the effects of mild conditions, easy operation, and low pollution

Inactive Publication Date: 2013-10-30
SHANGHAI INST OF PHARMA IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its preparation is mainly oxidized glutathione, the yield is about 58%, no relevant purity information is mentioned

Method used

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  • Method for separating and purifying reduced glutathione (GSH) from reduced glutathione contained fermentation leaching liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Take 2L of GSH extract, containing 12.8g of GSH. Adjust the extract to pH 4.0 and purify it with gel-type strong base anion exchange resin FPA40, wash the resin with 3 times the volume of the resin in deionized water, then desorb with 0.1mol / L hydrochloric acid, and collect the desorbed concentration higher than 1mg / ml liquid and concentrated to 200ml. Adjust the concentrated solution to pH 2.8 with halogen-containing non-polar adsorption resin SP207, wash the resin with 3 times the resin volume of deionized water, desorb with 2% ethanol aqueous solution, and collect the reduced form after comprehensively considering the influence of yield and purity. Glutathione concentration: the ratio (R / O) of oxidized glutathione concentration is greater than 20 desorption solution and concentrated to 30ml, stir at room temperature and add a small amount of seed crystals and then add 15ml ethanol, continue to stir until the mother liquor concentration Significantly lower again, sto...

Embodiment 2

[0039]Take 2L of GSH fermentation broth, containing GSH12.1g. Adjust the extract to pH 4.0, use gel-type strong base anion exchange resin FPA40, wash the resin with deionized water 3 times the volume of the resin, then desorb with 0.1mol / L hydrochloric acid, and collect the desorption solution with a concentration higher than 1mg / ml And concentrated to 200ml. Adjust the concentrated solution to pH 2.8 with halogen-containing non-polar adsorption resin SP207, wash the resin with 3 times the resin volume of deionized water, desorb with 2% ethanol, and collect reduced glutathione concentration: oxidized glutathione Concentrate the desorption liquid with a ratio of glycylidene concentration greater than 20 to 30ml, stir at room temperature and add a small amount of seed crystals, then add 15ml of ethanol, continue to stir until the concentration of the mother liquor no longer decreases significantly, stop stirring, filter, and dry the crystals to obtain crystals 5.9 g, observed u...

Embodiment 3

[0041] Take 2L of GSH extract, containing 12.1g of GSH and 11.3g of glutathione. Adjust to pH 4.0 and purify with macroporous strong base anion exchange resin IRA900, wash the resin with deionized water 3 times the volume of the resin, desorb with 0.1mol / L hydrochloric acid, collect the desorbed solution with a concentration higher than 1mg / ml and concentrate to 200ml. Adjust the pH of the concentrated solution to 2.8 and purify it with halogen-containing non-polar adsorption resin SP207, wash the resin with 3 times the volume of the resin in deionized water, desorb with 2% ethanol, collect the desorbed solution with an R / O ratio greater than 20 and concentrate to 25ml, stir at room temperature and add a small amount of seed crystals, then add 12.5ml of ethanol, continue to stir until the concentration of the mother liquor no longer significantly decreases, stop stirring, filter, and dry the crystals to obtain 5.0g of crystals, with a total yield of 41.3%. The weight of the so...

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Abstract

The invention provides a method for separating and purifying reduced glutathione (GSH) from a reduced GSH contained fermentation leaching liquid. The method comprises the following steps of: (1) treating the fermentation leaching liquid sequentially through anion exchange resin and a non-polar absorbent to obtain a reduced GSH contained solution; and (2) concentrating the reduced GSH contained solution, and then, crystallizing and drying. The reduced GSH separated and purified by using the method provided by the invention has the advantages that the purity of the reduced GSH can be up to over 98.5% (titration, drying), and relevant impurities conform to the requirement of European pharmacopoeia.

Description

technical field [0001] The invention relates to a method for separating and purifying reduced glutathione (abbreviated as GSH), in particular to a method for separating and purifying reduced GSH from a fermented liquid containing reduced GSH by a resin method. Background technique [0002] GSH is a tripeptide compound formed by condensation of glutamic acid, cysteine ​​and glycine through peptide bonds, and is a widely used active short peptide. The molecular weight of GSH is 307.33, the melting point is 189-193°C (decomposition), the isoelectric point is 5.93, and the crystal is colorless, transparent and slender rod. It is soluble in water, liquid ammonia, insoluble in alcohol, ether and acetone. GSH solid is relatively stable, and its aqueous solution is easily oxidized in air. The reduced structure is [0003] [0004] Since GSH is a very special amino acid derivative and a tripeptide containing a sulfhydryl group, it plays an important role in the living body. (1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/037C07K1/36C07K1/18C07K1/22
Inventor 赵波那可赵文杰许炜程晴华金媛媛潘兴荣赵云灿
Owner SHANGHAI INST OF PHARMA IND CO LTD
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