Process for producing concentrated and purified heparin

A production process, heparin technology, applied in the production process of concentrated and purified heparin, can solve the problems of low extraction rate of active ingredients, many operation steps, and large resin damage, and achieve the effects of shortening the production cycle, high investment efficiency, and low cost

Inactive Publication Date: 2010-09-15
XINJIANG LISHI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has low column efficiency, great damage to the resin, many operating steps, high cost, and low extraction rate of active ingredients.

Method used

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  • Process for producing concentrated and purified heparin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] a) Add about 50 g of water to 250 L of tissue homogenate of small intestinal mucosa of mammals such as pigs, cattle, sheep, etc., add NaCl to 3%, stir, stand still, and filter.

[0032] b) Add 0.25% trypsin to the filtrate, enzymatically hydrolyze at pH 8.5, 40°C for several hours, adjust the pH to 9.0 with NaOH, raise the temperature to 50°C, keep it warm for 2h, keep stirring, then quickly heat up to 92-95°C, keep for 10- After 15 minutes, filter with an 80-mesh filter while it is still hot, and use the filtrate (I) for later use.

[0033] c) Take an appropriate amount of strong alkaline D254 dry resin, soak it fully in distilled water, swell it and filter it dry, add an equal volume of ethanol or acetone and stir for 1 hour, wash it with distilled water and filter it dry, add 4 times the amount of 2mol / l hydrochloric acid solution Stir for 2 hours, wash with distilled water until neutral, filter dry; add 2 times the amount of 2mol / l sodium hydroxide solution and stir...

Embodiment 2

[0041] a) Dissolve 10 g of crude heparin in 100 ml of 3% sodium chloride solution, then adjust the pH to 9 with 2% sodium hydroxide solution, let stand at 18° C. for 6 hours, vacuum filter, and use the filtrate for later use.

[0042] b) Add 0.03% trypsin to the above liquid, enzymolyze at pH 8.5, 40°C for 4 hours, heat up to 98-100°C for 10 minutes, vacuum filter while hot, and filtrate (I) for later use.

[0043] c) Take an appropriate amount of strong alkaline D254 dry resin, soak it fully in distilled water, swell it and filter it dry, add an equal volume of ethanol or acetone and stir for 1 hour, wash it with distilled water and filter it dry, add 4 times the amount of 2mol / l hydrochloric acid solution Stir for 2 hours, wash with distilled water until neutral, filter dry; add 2 times the amount of 2mol / l sodium hydroxide solution and stir for 2 hours, wash with distilled water until neutral, filter dry; finally use 2 times the amount of 2mol / l hydrochloric acid solution S...

Embodiment 3

[0051] a) Add about 30 g of water to 250 L of tissue homogenate of liver, lung and other organs of mammals such as pigs, cattle, sheep, etc., add NaCl to 5%, stir, stand still, and filter.

[0052] b) Add 0.4% papain to the above liquid, enzymolyze at pH 8.5, 40°C for 4 hours, heat up to 98-100°C for 10 minutes, vacuum filter while hot, and filtrate (I) for later use.

[0053] c) Take an appropriate amount of strong alkaline D254 dry resin, soak it fully in distilled water, swell it and filter it dry, add an equal volume of ethanol or acetone and stir for 1 hour, wash it with distilled water and filter it dry, add 4 times the amount of 2mol / l hydrochloric acid solution Stir for 2 hours, wash with distilled water until neutral, filter dry; add 2 times the amount of 2mol / l sodium hydroxide solution and stir for 2 hours, wash with distilled water until neutral, filter dry; finally use 2 times the amount of 2mol / l hydrochloric acid solution Stir for 2 hours, wash with water until ...

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PUM

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Abstract

The invention discloses a process for producing concentrated and purified heparin, which comprises the following steps of: separating, concentrating and purifying heparin, impure protein, nucleic acid and rest sulfated polysaccharide at one time in solution of a coarse heparin product by using expansion column bed technology. The process changes the traditional process finished by multiple steps of a salt hydrolysis-ion exchange resin method. Fine heparin is separated, concentrated and purified by adopting the expansion column bed and ion exchange resin process, sedimentation is performed by adopting ethanol, dehydration is performed by adopting acetone, and the potency of the heparin is more than 180IU / mg. The expansion column bed and ion exchange resin process reduces the working procedure, shortens the time, lowers the cost, and improves the efficiency and quality.

Description

[technical field] [0001] The invention relates to a production process for concentrating and purifying heparin, in particular to a production process for concentrating and purifying heparin by using expanded column bed ion exchange adsorption chromatography technology. [Background technique] [0002] Heparin is an acidic mucopolysaccharide found in liver tissue when Mclean studied the coagulation mechanism in 1916. In 1939, Brinkhous et al. proved that heparin has anticoagulant activity. Since then, heparin has been valued by countries all over the world as a natural anticoagulant substance. Heparin is widely distributed in mammalian tissues, such as liver, lung, intestinal mucosa, heart, spleen, kidney, thymus, placenta, muscle and blood. Heparin is a glycosaminoglycan, which is a mixture of polysaccharide chains with different chain lengths composed of uronic acid and glucosamine. The molecular weight is between 3000-30000D, and the average molecular weight is about 12000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/10C12S3/02
Inventor 来海中赵文军卢耀勤严欢
Owner XINJIANG LISHI BIOTECH
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