Method for manufacturing high purity monosialogangliosides

Abstract

A method for preparing the Monosialotetrahexosylganglioside with high purity is provided, which comprises the following process steps: (1) the fresh brain tissue of the licestock is used to prepare the acetone powder; (2) the acetone powder prepared in the step (1) is dissolved by the extracting solution and stirred for 4h to 5h, and is separated centrifugally and collects the supernatant, the supernatant is purified by LV chromatographic column to gain the total ganglioside; (3) the total ganglioside prepared in the step (2) is separated by MonoQ column chromatography to gain the ganglioside GM1 with purity of 90 per cent to 95 per cent; (4) the GM1 prepared in the step (3) is separated by the hydrophobic chromatography to gain the ganglioside GM1 with purity more than 98 per cent; (5) the GM1 aqueous solution prepared in the step (4) is hyperfiltrated by the hyperfiltration membrane with 50,000 to 100,000 MWCO, and the gained filter liquor produces the GM1 dry powder through freezing and drying. The method not only has simple process flow and safe operation, but also can improve the yield of the GM1 and avoid the pollution of other phospholipids.

Description

technical field [0001] The invention belongs to the field of preparation of monosialotetrahexosyl ganglioside, and in particular relates to a method for preparing monosialotetrahexosyl ganglioside. Background technique [0002] Monosialotetrahexose ganglioside (monosialotetrahexose ganglioside, referred to as GM1, "GM1" in the description below and in the claims is "monosialotetrahexose ganglioside") is the only currently known penetrating Gangliosides of the blood-brain barrier. The molecular weight of GM1 is 1547, and it is mainly located in myelin sheath, neuron cell membrane and axon, so it is abundant in the brain (about 730 nmol GM1 per gram of brain tissue), and GM1 is also contained in peripheral nerves such as sciatic nerve and femoral nerve. After the central and peripheral nerves are damaged, endogenous GM1 is insufficient, and exogenous GM1 gathers in the damaged area, integrates into the plasma membrane of the cell, and becomes a component of the cell through e...

AI Technical Summary

Problems solved by technology

The weak point of this kind method is: 1, the flow process of the initial extract of mixed gangliosides is more complicated. In this step, the filtrate obtained by ultrafiltration membrane ultrafiltration is desalted, concentrated and then freeze-dried to easily produce explosions. Potential safety hazard. In addition, the formula of the extract in this step hinders the yield of increasing ganglioside GM1

Although this method can obtain GM1 dry powder with a purity greater than 98%, it has the following problems: 1. The process of extracting the total ganglioside step is more complicated

2. Dissolve the concentrated hydrolyzate in an organic solvent mixture after desalination for anion exchange column chromatography, in which the ion exchange resin has no selectivity for GM1, but preferentially adsorbs substances with low polarity, so although water-soluble impurities can be separated and removed , but the phospholipids that also have hydrophilic groups and hydrophobic groups are difficult to remove, so phospholipids are highly polluted and contain high phosphorus

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