How does the ceramic membrane remove impurities from the crude heparin sodium?

How does the ceramic membrane remove impurities from the crude heparin sodium?

Heparin is a glycosaminoglycan widely found in animal organs and small intestinal mucosa, lung and other tissues. In the conventional extraction process, a large amount of impurities such as proteins, peptides, amino acids and nucleic acids affect the purification process of heparin sodium. The content of these impurities in crude heparin sodium directly affects the activity and product quality of heparin sodium products.

Traditional methods to remove macromolecular proteins, peptides, amino acids, nucleic acids and other impurities in enzymatic hydrolysates. Currently, most methods are used in China: one is to use traditional filtration methods such as plate and frame filtration, drum rotation, centrifugal separation or separation equipment. This filtration method can only filter out visible impurities and a small amount of large particle impurities, and there are a large number of water-soluble protein impurities which are difficult to be filtered out in the enzymatic hydrolysate; the other is flocculation and sedimentation by adding a complexing agent (for example, Addition of complexing agents, alum, calcium carbonate, polyaluminum chloride, etc.) or isoelectric precipitation, such a method can make complexing agents and protein complexes to synthesize larger particles, and then through the plate frame filtration, centrifugation Filtering and other traditional "dead end filtration" and "filter cake filtration" and other filtration methods are removed. The disadvantages of this method are the amount of complex addition, the concentration of the complex, the speed and cycle of adding the complexing agent, and the need for a certain static. At the same time, this complexing agent is easy to cause heparin activity loss due to a certain complexation with heparin sodium. The above-mentioned conventional processes have been used for a long time and are generally used. However, these conventional processes can only crudely separate solids such as large suspended solids in the enzymatic hydrolysate, and cannot dissolve the soluble proteins, impurities, pigments, etc. which are abundantly present in the enzymatic hydrolysate. Separation.

specific contents

A method for removing impurities in the extraction process of crude heparin sodium by using a ceramic membrane is provided to overcome the disadvantages of the prior art.

For the purpose of the present invention, such a method for removing impurities in crude heparin sodium using a ceramic membrane is characterized by comprising the following process steps:

The small intestinal mucosa and water are mixed at a mass ratio of 1:6-8, followed by adding NaCl and protease, then heating at 50-60 ° C, and then digesting for 3 to 5 hours with stirring. The amount of NaCl added is pig small intestine. 1/8~1/6 of the mass of the mucous membrane, the protease is added in an amount of 0.2-0.4% of the mass of the pig small intestine;

The obtained enzymatic hydrolysate is filtered and filtered through a 5 nm ceramic membrane at 50 ° C to 60 ° C and 0.2 to 0.3 MPa;

Adding an ion exchange resin to the filtered clear liquid, and stirring and adsorbing for 6 to 8 hours;

The ion exchange resin adsorbing heparin sodium is washed with 4~7% NaCl aqueous solution, and then the ion exchange resin is eluted with 20~24% NaCl aqueous solution to obtain an eluate containing heparin sodium;

85~95% ethanol is added to the eluate obtained above. When the concentration of ethanol in the solution reaches 40-50% (w/v), the precipitate is allowed to stand for 2 hours, then dehydrated and dried to obtain crude heparin sodium. The amount added is 2 to 4 times the mass of the eluent.

skill improved:

Sealed at room temperature to ensure product activity and meet current GMP certification requirements

The enzymatic hydrolysate of the invention completes the separation process of materials in the closed pipeline and the membrane cavity, and the solution is not directly in contact with the atmosphere, and meets the requirements of GMP production.

High separation accuracy and high filtration efficiency

The pore size distribution of the inorganic ceramic membrane layer is very narrow, and the absolute precision of the 5 nm pore size ceramic membrane is high, which can ensure that a large amount of impurities in the enzymatic hydrolysate cannot easily penetrate the membrane layer, and the rejection rate of impurities is over 99%, in the enzymatic hydrolysate The retention rate of macromolecular proteins is much higher than that of traditional filtration and complexing agent sedimentation.

Can greatly improve the yield of wide products

The active ingredient in the enzymatic hydrolysate can be collected to the maximum extent, and the yield of the product can be improved. Compared with the existing conventional filter bag, filter cloth and plate and frame filtration method, the yield can reach 99.5%.

Guaranteed crude heparin sodium quality

Since the content of a small amount of small molecular proteins in the filtrate is significantly reduced compared with the conventional method, and the permeate is clear and transparent, the stability of the product quality is ensured.

Wastewater discharge is greatly reduced

The ceramic membrane filter removes the impurities in the solution, and compared with the wastewater discharge pollution in the traditional process, the sewage discharge can be minimized, the COD in the wastewater is significantly reduced, the sewage treatment cost is effectively reduced, and the environmental protection pressure is alleviated.

The filtrate is clear and transparent, which reduces the difficulty of subsequent adsorption treatment

The ceramic membrane filtrate greatly simplifies the subsequent ion exchange impurity removal step and reduces the contamination degree of the subsequent ion exchange resin, thereby prolonging the service life of the ion exchange resin, and the service life of the ion exchange resin can be extended by 40 to 50%, thereby saving the consumption cost of the resin auxiliary material.

No need to add additives

Since the membrane filtration process is a simple physical filtration process, the material components are separated by a high-precision membrane pore size, and no additional additives are required for flocculation. The filter residue after filtration is free from pollution and contains.