Preparation method of monosialoteterahexosylganglioside

A technology of ganglioside and monosialic acid, which is applied in the field of medicine and biology, can solve the problems of low yield and purity of GM1, large amount of organic solvent, and difficult waste disposal, etc., and achieves the reduction of water-soluble impurities, high recovery rate, and reduction of The effect of residual impurities

Active Publication Date: 2012-10-17
四川奇格曼药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] So far, in the extraction and purification of GM1, this kind of process often has many deficiencies, which are not conducive to the development of industrial prod

Method used

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  • Preparation method of monosialoteterahexosylganglioside
  • Preparation method of monosialoteterahexosylganglioside
  • Preparation method of monosialoteterahexosylganglioside

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 pig brain selection and pretreatment

[0043] Select fresh pig brains or frozen pig brains that have passed the animal epidemic prevention inspection, remove meninges and blood vessels, wash away blood stains with deionized water, rinse 3 times, mince, and homogenate. After pulverization, use acetone equivalent to 5 times the volume of the pig brain used in the test, stir for 15 minutes, and stand at room temperature for immersion treatment for 12 hours. Filter and collect the filter residue. The filter residue was moved to a distillation pot for distillation and drying, and the dry residue was collected to obtain porcine brain acetone powder, which was sealed and subpackaged, and stored for later use.

Embodiment 2

[0044] Example 2 Extraction of total gangliosides

[0045] Pump 500L of extraction buffer into the stirring tank, then add 100 kg of pig brain acetone powder, stir and extract for 60 minutes, filter, and collect the precipitate and filtrate. The extraction buffer consists of chloroform, methanol and water in a volume ratio of 4:8: 3 ratio mix. Add 500L of the extraction buffer to the precipitate, stir and extract for 60 minutes, filter, and collect the precipitate and extract. Combine the filtrate and the extract, add water equivalent to 20% of the volume of the extract to the extract, and mix well to obtain a mixed solution; leave the mixed solution at room temperature for 12 hours to separate layers, and take the supernatant. Add NaOH powder to the supernatant so that the mass fraction of NaOH in the supernatant is 0.3%, stir and dissolve, the pH value reaches 10.0-11.0, saponify at 60° C. for 45 minutes, and cool the saponified solution for later use.

[0046] After the p...

Embodiment 3

[0047] The extraction of embodiment 3 GM1 crude product

[0048] Add 0.2% hydrochloric acid (that is, 120 ml) to 60 L of the concentrated ganglioside prepared in Example 2 to adjust the pH to 2.5 to obtain a mixed solution. Stir the reaction at 80°C for 2 hours, cool to room temperature, and set aside. Add acetone equivalent to 5 times the volume of the mixed solution, and cryoprecipitate at -10°C for 12 hours. Centrifuge, collect the precipitate by filtration with a 200-mesh filter cloth, and obtain a crude product containing GM1, which contains a mass fraction of 35% "crude product 1".

[0049] Purify "crude product 1": take 300 kilograms of crude silica gel, and use mobile phase chloroform / methanol / water (the volume ratio of chloroform, methanol and water is 45:40:5) to wet-pack to a 80×100cm chromatographic column middle. Fully dissolve "crude product 1" with 80L of mobile phase per 10kg, and load the sample at a flow rate of 50L / hour. Elute with the above-mentioned mo...

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Abstract

The invention discloses a method for preparing high-purity monosialoteterahexosylganglioside (GM1), comprising the steps of preparing acetone powder of pig brains, preparing total ganglioside and conducting acid hydrolysis. According to the invention, the yield and purity of the obtained monosialoteterahexosylganglioside are raised, the application amount of organic solvent is saved, the extraction time is saved, the efficiency is raised, and convenience is brought to the subsequent process.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and in particular relates to a method for preparing high-purity monosialotetrahexosyl ganglioside (GM1) by using fresh pig brain as a raw material. Background technique [0002] Monosialotetrahexosylganglioside (GM1) is a glycosphingolipid substance containing sialic acid, and is the only known ganglioside that can penetrate the blood-brain barrier. GM1 was first discovered by Klenk in Germany. It is widely present in the cell membranes of mammals, especially in the nervous system. It is embedded in the double-layered lipid membrane structure of cells. 10% of the quality. GM1 plays an essential role in the occurrence, growth, differentiation and regeneration of the human nervous system. Years of experiments and clinical reports have shown that GM1 can promote the recovery of damaged nerves, has the effect of promoting "neural remodeling", and is one of the main substances involved ...

Claims

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Application Information

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IPC IPC(8): C07H15/10C07H1/08
Inventor 黄洪涛张宪姚静石延宾李豫俭杨琴飞胡伟魏勇
Owner 四川奇格曼药业有限公司
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