Study On Extraction And Purification Of Heparin Sodium

In this paper,the technology of preparing heparin sodium by enzymatic hydrolysis and membrane separation was studied: first,the aqueous solution of porcine intestinal mucosa under alkaline conditions was used to obtain heparin sodium extract,and after coarse filtration,heparin sodium was collected by resin adsorption process to remove some impurities,then the heparin sodium was washed off by resin elution,Then,the heparin sodium elution solution was purified by membrane separation technology,and the heparin sodium concentrate containing little impurities was obtained,and then the heparin sodium was obtained by precipitation and drying the concentrate solution.Based on actual production,this paper provides theoretical basis for industrialized production design.Porcine intestinal mucosa by enzymatic hydrolysis using an aqueous solution of alkaline protease in the solid-liquid ratio 1:25 conditions,the temperature during the hydrolysis Effects,time,pH,salt concentration and other factors of the results using orthogonal design software the key factor for optimal analysis,the best explore hydrolysis conditions;different types of basic anion adsorption resin was subjected to hydrolysis,and the adsorption rate and adsorption rate of the resin for comparison of different heparin types of basic anion adsorption,then most studies suitable resin adsorption adsorption process temperature,pH and salt concentration,to explore the best adsorption process parameters;Research microfiltration membrane,ultrafiltration membrane and 2000 D ultrafiltration eluate 40000 D protein,P-containing impurities and sodium heparin rejection after washing the membrane and the recovery rate,determine the feasibility of a membrane separation technology impurity;on quality of heparin obtained new process and traditional process system,results were as follows:1.The best technical parameters of enzymatic hydrolysis of alkaline protease are: the material liquid ratio is 1:25,the enzyme substrate is 0.8%,the enzymatic hydrolysis temperature is 55 ?,the enzymatic hydrolysis time is 3.0h,the enzymatic hydrolysis is pH9.0,and the enzymatic hydrolysis salt concentration 2.5°bé.2.The D208 type alkaline anion adsorption resin adsorption process the best technical parameters: Adsorption time 8h,adsorption temperature 50 ?,adsorption pH8.5,adsorption salt concentration 2.5°bé.3.At the liquid temperature of 40 ?,ph 8.0,operating pressure of 0.25 MPa under the condition of the filter membrane C protein interception rate of 52.3%,containing P material interception rate of 54.9%,heparin sodium interception rate 98.1%;at room temperature,Under the condition of ph 8.0 and operating pressure of 1.0MPa,the protein interception rate of 40000 D ultrafiltration membrane was 62.9%,with P impurity interception rate 70.2%,and heparin sodium loss rate 4.7%;at room temperature,ph 8.0,operating pressure of 1.25 MPa under the conditions of 2000 D ultrafiltration membrane protein removal of 57.9%,containing p impurities removed 67.2%,heparin sodium loss rate of 1.1%,the liquid concentration of 12.4 times.4.The new process made heparin sodium titer of 186.3u/mg,protein impurity 0.21%,the traditional process of heparin sodium titer of 197.2u/mg,protein impurity 0.13%,and the two processes prepared heparin sodium solution clarity and color,nucleic acid impurities are in line with the standard.The loss rate of heparin sodium was 28.35% in the process of the new process,but the loss rate of the traditional process was about 55.38%,and the new process had a 36.9% higher yield than the traditional process of heparin sodium.