Production and purification process of high-purity low-molecular-weight heparin sodium

Abstract

The invention discloses a production and purification process of high-purity low-molecular-weight heparin sodium, and the proocess comprises the following steps of: S1, weighing 80-120g of crude heparin sodium, performing dissolving with purified water, controlling the temperature of the solution to be below 30DEG C, and conducting stirring for dissolving to prepare a 10%-20% (w / v) crude heparin sodium solution; S2, adding the crude heparin sodium solution in a NaCl solution with a concentration of 2% to prepare a 5%-10% (w / v) crude heparin sodium solution, adding alkaline protease, and carrying out heat preservation enzymolysis at 35-50DEG C for 3-6h to obtain an enzymatic hydrolysate; and S3, carrying out centrifugal separation on the enzymatic hydrolysate obtained in S2 to obtain a supernatant and a precipitate, and discarding insoluble impurities. According to the method, macromolecular substances such as residual protein in the produced low-molecular-weight heparin are interceptedby using an ultrafiltration method, so that the safety of the low-molecular-weight heparin is improved, and the low-molecular-weight heparin sodium is low in protein content, high in purity and highin yield.

Description

technical field [0001] The invention relates to the field of biochemical technology, in particular to a production and purification process of high-purity low-molecular-weight heparin sodium. Background technique [0002] Heparin has been the drug of choice for the prevention and treatment of thrombosis since the 1980s. Low-molecular-weight heparin is made from unfractionated heparin through specific chemical cutting and purification. Due to its definite curative effect, small and predictable side effects, it has gradually replaced the market position of traditional heparin. [0003] Existing purification methods are mainly divided into the following types: (1) decolorize enoxaparin sodium with activated carbon, filter through macroporous resin, and freeze-dry the filtrate to obtain purified enoxaparin sodium. The decolorization effect of this method is better, but the disadvantages are There will be a large amount of products adsorbed in activated carbon and resin, and the...

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Problems solved by technology

[0003] Existing purification methods are mainly divided into the following types: (1) decolorize enoxaparin sodium with activated carbon, filter through macroporous resin, and freeze-dry the filtrate to obtain purified enoxaparin sodium. The decolorization effect of this method is better, but the disadvantages are There will be a large amount of products adsorbed in activated carbon and resin, and the yield can only reach about 60%; (2) adjust the pH of enoxaparin sodium with sodium hydroxide, decolorize with hydrogen peroxide, and freeze-dry after washing with sodium chloride and ethanol , to obtain purified enoxaparin sodium, this method requires the use of highly oxidizing hydrogen peroxide, which will easily lead to the generation of other by-products while removing the pigment, and it will pollute the environment more, and it is difficult to meet the requirements of the environmental impact assessment; (3) will Enoxaparin sodium passes through hydrophobic chromatography column and anion exchange resin in turn, after desalination by nanofiltration, precipitation and drying to obtain purified enoxaparin sodium. This method requires multiple chromatography columns, and the product loss is very large; (4) Enoxaparin sodium is passed through filtration, microfiltration and ultrafiltration devices in sequence, and purified enoxaparin sodium is obtained after freeze-drying. The product loss of this method is small, but the operating pressure of this graded ultrafiltration is relatively high, which is not suitable for industrial use. Application; (5) enoxaparin sodium is repeatedly treated with sodium chloride solution and methanol to finally obtain enoxaparin sodium with up to standard clarity. This method is easy to operate and easy to realize in industry, but its purification effect is relatively poor, and It is difficult to accurately determine the number of repeated treatments each time, which is not conducive to the establishment of production standards