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Coproduction process of pig brain protein hydrolysate and monosialoganglioside

A technology for ganglioside and hydrolyzed protein, which is applied in the field of co-production technology of porcine brain hydrolyzed protein and monosialotetrahexosylganglioside

Active Publication Date: 2011-06-15
SHANDONG NEWTIME PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the separation and purification of GM1, there are many reports at home and abroad. Chinese patent CN1158295C discloses a method for preparing GM1 with ultrafiltration membrane combined with affinity chromatography or high performance liquid phase. Although the purity of GM1 obtained in this invention is high, the equipment used High requirements, large investment, and small output, not suitable for industrial production
Chinese patent CN1400216A discloses a process of selecting a suitable organic solvent for extraction according to the properties of gangliosides, thereby increasing the yield and purity of GM1, but the purity of GM1 obtained by this invention is only 81%, which is far from the standard for pharmaceutical injections
For the separation and purification of cerebroprotein hydrolyzate, Chinese patent CN101524370 discloses a preparation process of pig cerebroprotein hydrolyzate, but the cerebroprotein hydrolyzate prepared by this process can only meet the requirements of edible or oral medicine, and has not yet reached the level of medical injection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Take 100kg of fresh pig brain, remove the capsule and blood vessels on the surface, pulverize it with a 120-mesh colloid mill, add 1 times the volume of absolute ethanol, homogenize at room temperature for 4 hours, and filter under reduced pressure to collect the filter cakes respectively and ethanol filtrate;

[0020] (2) Add 1 times the volume of ethyl acetate and 0.5 times the volume of purified water to the filtrate of step (1), at 35°C, stir for 30 minutes and then stand to separate layers, remove the lower ethanol aqueous phase, and add purified water Adjust the ethanol concentration to 60%, load the non-polar macroporous adsorption resin, wash with purified water for 2 times the column volume after the adsorption is saturated, elute with 80% ethanol solution, and collect the eluate. When concentrated under reduced pressure to 1 / 4 of the original volume at 60°C, 2 times the volume of pre-cooled acetone was added, precipitated for 6 hours, filtered under reduce...

Embodiment 2

[0027] (1) Take 100kg of fresh pig brain, remove the capsule and blood vessels on the surface, pulverize it with a 120-mesh colloid mill, add 4 times the volume of absolute ethanol, homogenate at room temperature for 2 hours, and filter under reduced pressure to collect the filter cakes respectively and ethanol filtrate;

[0028] (2) Add 0.5 times the volume of ethyl acetate and 0.1 times the volume of purified water to the filtrate of step (1), at 30°C, stir for 30 minutes and then stand for stratification, remove the lower ethanol water phase, and add purified water Adjust the ethanol concentration to 30%, load the sample with non-polar macroporous adsorption resin, wash with purified water for 2 times the column volume after the adsorption is saturated, elute with 80% ethanol solution, and collect the eluate. Concentrate under reduced pressure to 1 / 4 of the original volume at 57°C, add 1 volume of pre-cooled acetone, precipitate for 10 hours, filter under reduced pressure a...

Embodiment 3

[0035] (1) Take 100kg of fresh pig brain, remove the capsule and blood vessels on the surface, pulverize it with a 120-mesh colloid mill, add 2 times the volume of absolute ethanol, homogenate at room temperature for 3 hours, and filter under reduced pressure to collect the filter cakes respectively and ethanol filtrate;

[0036] (2) Add 1 times the volume of ethyl acetate and 0.1 times the volume of purified water to the filtrate of step (1), stir at 40°C for 30 minutes, then let stand to separate layers, remove the lower ethanol aqueous phase, and add purified water Adjust the ethanol concentration to 50%, load the non-polar macroporous adsorption resin, wash with purified water for 2 times the column volume after the adsorption is saturated, elute with 80% ethanol solution, and collect the eluate. When concentrated under reduced pressure to 1 / 4 of the original volume at 55 ° C, 2 times the volume of pre-cooled acetone was added, precipitated for 5 hours, filtered under reduce...

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PUM

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Abstract

The invention discloses a method for obtaining pig brain protein hydrolysate and monosialoganglioside (GM1) from fresh pig brain through fractional extraction. The main process routes are as follows: (1) firstly, adding polar organic solvent homogenate fresh pig brain, and respectively collecting a filter cake and filtrate after filtration; (2) adding a certain amount of low-pole organic solvent and purified water in the filtrate from the step (1) for Folch lamination, then respectively carrying out resin column chromatography, hydrolysis and silicagel column chromatography so as to obtain high pure monosialoganglioside; and (3) adding a certain amount of acetone in the filter cake from the step (1) for degrease, filtering and drying, adding salt buffer solution and hydrolase for hydrolysis, and carrying out inacitivation, ultrafiltration purification, ultrafiltration concentration and vacuum freeze drying so as to obtain medical injection-stage pig brain protein hydrolysate.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and in particular relates to a co-production process of pig brain hydrolyzed protein and monosialotetrahexosyl ganglioside. Background technique [0002] Monosialotetrahexosyl ganglioside (GM1) is a membrane glycolipid containing N-acetylneuraminic acid (sialic acid), which is extracted and refined from fresh pig brain. It can repair nerve damage and enhance human brain Functional development, has good therapeutic and health care effects on nervous system diseases. For the separation and purification of GM1, there are many reports at home and abroad. Chinese patent CN1158295C discloses a method for preparing GM1 with ultrafiltration membrane combined with affinity chromatography or high performance liquid phase. Although the purity of GM1 obtained in this invention is high, the equipment used The requirements are high, the investment is large, and the output is small, so it is not suitable for ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H15/10C07H1/08C12P21/04A61K38/01A61K35/30A61P25/00
Inventor 赵志全董惠钧
Owner SHANDONG NEWTIME PHARMA
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