Preparation of kilogram-grade scale high-purity monosialotetrahexosylganglioside

A ganglioside and monosialic acid technology, applied in the field of biomedicine, can solve the problems of difficulty in extraction and separation of high-purity GM1, no reports and applications, and difficulty in large-scale clinical application. Increased yield and production efficiency, low product cost

Active Publication Date: 2008-05-14
LUNAN PHARMA GROUP CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many types of gangliosides, and their content in brain tissue is small, especially their molecular composition, structure, physical and chemical properties and other factors are very similar, so the extraction and separation of high-purity GM1 is difficult, especially in large-scale industrial extraction processes. No reports and applications
So far, most domestic research on GM1 extraction and separation is still at the laboratory scale and level, and h

Method used

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  • Preparation of kilogram-grade scale high-purity monosialotetrahexosylganglioside

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of high-purity GM1 freeze-dried powder

[0024] 1) Centrifuge the fresh pig brain tissue at 5000rpm to remove free water, pulverize it with a colloid mill, homogenize it at a high speed of 3000r / min for 30min, and pour it into the extraction tank and add 10 times the volume of methanol water (2:0.8) solution , stirred at a low temperature of 2°C for 12 hours, and extracted;

[0025] 2) Use a plate and frame filter to achieve solid-liquid separation of the extract, and then apply it to a macroporous adsorption resin D-101 chromatographic column, first wash off impurities with low-concentration methanol water (1:2), and then elute with anhydrous methanol Collecting the eluate containing gangliosides;

[0026] 3) The ganglioside eluate was concentrated in vacuum at 40°C and freeze-dried at -20°C, dissolved in 0.5M NaCl solution and adjusted to pH 3.5 with glacial acetic acid, hydrolyzed at 60°C for 3 hours, and used continuously during the hydrolysis...

Embodiment 2

[0030] Example 2 Preparation of high-purity GM1 freeze-dried powder

[0031]1) Centrifuge the fresh pig brain tissue at 5000rpm to remove free water, pulverize it with a colloid mill, homogenize it at a high speed of 3000r / min for 30min, and pour it into the extraction tank and add 10 times the volume of methanol water (2:1) solution , stirred at a low temperature of 8°C for 24 hours, and extracted;

[0032] 2) Use a plate and frame filter to achieve solid-liquid separation of the extract, and then apply it to a macroporous adsorption resin D-101 chromatographic column, first wash off impurities with low-concentration methanol water (1:3), and then elute with anhydrous methanol Collecting the eluate containing gangliosides;

[0033] 3) The ganglioside eluate was concentrated in vacuum at 80°C and freeze-dried at -60°C, dissolved in 0.5M NaCl solution and adjusted to pH 4.0 with glacial acetic acid, hydrolyzed at 75°C for 5 hours, and used continuously during the hydrolysis pr...

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Abstract

The invention discloses a manufacturing technique for extracting, separating and purifying kilogram-grade scale high purity GM1 using fresh pig brain tissue as raw material, which is characterized in that large sized extraction pot and adsorption resin with large holes, DEAE Sephadex A-25 and silica gel layer are utilized to build a set of manufacturing technique. The invention has the advantages of elimination of toxic solvent chloroform in mobile phase in traditional method, less pollution for whole process, simple process, low cost and applicability to industrialized production.

Description

Technical field [0001] The invention relates to a preparation method of kilogram-scale high-purity monosialotetrahexosyl ganglioside (GM1), which belongs to the field of biomedicine. Background technique [0002] Gangliosides (GLS) are a class of acidic glycolipids containing sialic acid, and are important components of mammalian cell membranes, with the highest content in brain tissue. The composition of gangliosides is very complex. Different gangliosides contain different numbers of sialic acids and connection methods of sugar groups. G can be used to represent gangliosides, and M, D, and T can represent single, di, and tri Sialic acid group, the subscript numbers 1, 2, and 3 represent tetraglycosyl, triosyl, and diglycosylceramide respectively, and GM1 represents a ganglioside composed of a single sialic acid group and tetraglycosylceramide. . Studies have shown that gangliosides with significant biological functions are GM1 connected with a single sialic acid, which c...

Claims

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Application Information

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IPC IPC(8): C07H15/10A61K35/30
Inventor 赵志全张贵民董惠钧强红刚
Owner LUNAN PHARMA GROUP CORPORATION
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