Method for rapidly and efficiently preparing high-purity ganglioside

Abstract

The invention relates to the field of biopharmaceuticals, in particular to a method for rapidly and efficiently preparing high-purity ganglioside. The method comprises the following specific steps ofextraction of a pig brain extract, GM1 silica gel chromatography purification and cationic resin purification, wherein the extraction of the pig brain extract comprises the following steps of extraction, stratification, saponification, resin chromatography, acid hydrolysis, acetone precipitation filtration and drying. Through extraction of the ganglioside with a heat extraction method, the extraction efficiency is improved greatly. The GM1 content is increased by more than twice through acid hydrolysis. Moreover, a high-pressure system is used for performing silica gel purification, thereby greatly shortening the purification time while improving the separation efficiency. Rapid and effective separation of GM1 and impurities is realized, on-line detection can be realized, subsequent point-plate detection is not needed, and the production efficiency is greatly improved. Moreover, substances affecting the clarity of an injection are removed through cationic resin chromatography, so thatthe occurrence rate of adverse reactions after treatment is reduced greatly.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method for rapidly and efficiently preparing high-purity gangliosides. Background technique [0002] Gangliosides are the most complex glycosphingolipids and exist in trace amounts in almost all tissues of vertebrates, but the highest content in mammalian brain and spinal cord tissues is about 0.3%. At present, the purification of GM1 is mainly based on atmospheric pressure silica gel column. For example, a Chinese patent discloses a method for extracting and separating gangliosides, application number: 201510598064.0, application date: [0003] 2015.09.18, it includes three steps: 1. The mammalian nerve tissue is placed in an alcohol solvent for heating, stirring and refluxing extraction, and then the extract is stirred and cooled to obtain a precipitate I containing gangliosides by filtration; 2. Precipitation The substance I is heated and dissolved in the organic solvent I...

AI Technical Summary

Problems solved by technology

[0003] 2015.09.18, it includes three steps: 1. Place the mammalian nervous tissue in an alcohol solvent, heat, stir and reflux to extract, then cool down the extract by stirring, and filter to obtain the precipitate I containing ganglioside components; 2. Precipitate Heat and dissolve substance I in organic solvent I, and maintain the heating temperature to carry out impurity adsorption under stirring conditions, filter the resulting solution and concentrate it with acetone for low-temperature precipitation, and filter to obtain precipitate II enriched in ganglioside components; 3. Precipitation Compound II is heated and stirred with organic solvent II and washed, and the precipitate is ganglioside by filtration. This method has relatively low cost, but has the problems of long production cycle and low yield, and there is an urgent need for a method with fast extraction speed, high extraction purity and production efficiency. Efficient method for preparing gangliosides