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Technology for extracting heparin sodium from intestinal mucosa with trypsin method

A technology of trypsin and intestinal mucosa, which is applied in the field of extracting heparin sodium from intestinal mucosa by trypsin method, can solve the problems of long production cycle, low product purity, environmental pollution, etc., and achieves reduction of odor, high product purity, and improved naturalness. Effect

Inactive Publication Date: 2013-07-03
HANGZHOU LONGYANG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The defects of the traditional enzymatic hydrolysis production method of heparin sodium are: long production cycle, high energy consumption, low yield, the mucous membranes of 2800-3000 pig small intestines are needed to produce 100 million international units of heparin sodium crude product, the product purity is low, and the yield of heparin sodium is low. Potency is at most 80U / mg, and produces a large amount of intestinal residue and waste water, polluting the environment
2709 protease is a synthetic product, which reduces the naturalness of heparin sodium products, and the use of 2709 protease for enzymatic hydrolysis produces a large odor during the production process and pollutes the environment
Simultaneously the product yield of this method, purity are also lower

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Trypsin enzyme solution preparation: Take fresh porcine pancreas, remove residual grease on the surface, grind it with a meat grinder, and mix the ground porcine pancreas:saturated lime water=1:10 by weight to obtain trypsin enzyme liquid.

[0042] (2) Enzymolysis: Add 1000L of porcine small intestine mucosal slurry (obtained from pig small intestines treated by an intestinal scraper) into the enzymatic hydrolysis tank. The porcine small intestinal mucosal slurry is a mixture of porcine small intestinal mucosa: water = 1: 3 by weight, and add salt to the enzymatic hydrolysis tank Sodium chloride solution with a temperature of 22° adjusts the salinity of the feed liquid in the enzymolysis tank to 5±0.5°, and controls the pH of the feed liquid at about 8.5; then add 15kg of enzyme to the enzymolysis tank for every 1000L feed liquid For the trypsin enzyme solution, the temperature of the feed solution is controlled at 53°C, the salinity is 4.2±0.2°, and the pH is abo...

Embodiment 2

[0054] (1) Trypsin enzyme solution preparation: Take fresh porcine pancreas, remove residual grease on the surface, grind it with a meat grinder, and mix the ground porcine pancreas:saturated lime water=1:9 weight ratio evenly to obtain trypsin enzyme liquid.

[0055] (2) Enzymolysis: Add 2000L of porcine small intestinal mucosal slurry into the enzymatic hydrolysis tank. The porcine small intestinal mucosal slurry is a mixture of porcine small intestinal mucosa: water = 1: 2.5 by weight. Add sodium chloride solution with a salinity of 21° to the enzymatic hydrolysis tank to adjust the enzymatic hydrolysis tank The salinity of the feed liquid is 5±0.5°, and the pH of the feed liquid is controlled at 8.0; then, add 10kg of enzyme per 1000L feed liquid to the enzymolysis tank, and control the temperature of the feed liquid at 50°C. Salinity is 4.2±0.2°, pH 6.5 is kept for 3.5 hours; finally, the temperature is raised to 85°C and kept for 25 minutes to obtain the enzymatic hydr...

Embodiment 3

[0066] (1) Trypsin enzyme solution preparation: Take fresh porcine pancreas, remove residual grease on the surface, grind it with a meat grinder, and mix the ground porcine pancreas:saturated lime water=1:12 by weight evenly to obtain trypsin enzyme liquid.

[0067] (2) Enzymolysis: Add the porcine small intestinal mucosal slurry into the enzymatic hydrolysis tank, which is a mixture of porcine small intestinal mucosa: water = 1:4 by weight, and add sodium chloride solution with a salinity of 24° to the enzymatic hydrolysis tank to adjust the enzymatic hydrolysis tank The salinity of the feed liquid is 5±0.5°, and the pH of the feed liquid is controlled at 9.0; then add 20kg of enzyme per 1000L of feed liquid to the enzymolysis tank, and the temperature of the feed liquid is controlled at 56°C. Temperature 4.2±0.2°, pH 7.5 and keep for 2.5 hours; finally raise the temperature to 88°C, keep for 20min to obtain the enzymatic hydrolysis solution, filter to remove the residue. ...

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Abstract

The invention relates to the technical field of heparin sodium production, and provides a technology for extracting heparin sodium from intestinal mucosa with a trypsin method. The technology is characterized by adopting a fresh pig pancreas as the raw material to prepare a trypsin liquid and to extract the heparin sodium, therefore, utilization of artificial compositions is reduced greatly, and the product nature is improved; furthermore, as the trypsin liquid is adopted to extract the heparin sodium, peculiar smell in a production process can be reduced to a large extent, and the environmental protection is facilitated. According to the invention, the fresh pig pancreas is adopted as the raw material to prepare the trypsin liquid and to extract the heparin sodium, the yield is high, only about 1600 pig small intestines are needed to produce hundred million international units of the crude heparin sodium product, the product purity is high, and the titer of the heparin sodium is larger than 100 U / mg.

Description

technical field [0001] The invention relates to the technical field of heparin sodium production, in particular to a technique for extracting heparin sodium from intestinal mucosa by trypsin method. Background technique [0002] Heparin sodium, also known as heparin, is a natural anticoagulant substance containing acidic mucopolysaccharides containing sulfate groups. Heparin sodium is white or off-white powder, odorless, tasteless, hygroscopic, soluble in water, insoluble in organic solvents such as ethanol, acetone, and dioxane. [0003] Heparin sodium widely exists in mammalian liver, lung, and intestinal mucosa, and mostly exists in complexes combined with proteins. Enzyme hydrolyzed protein can separate heparin sodium, which is mucopolysaccharide containing sulfuric acid, amino group and aldonic acid. At pH 8-9, it is negatively charged, and can perform ion exchange with anion exchangers for rough separation, and the polysaccharide solution is precipitated in high-conc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/10
Inventor 花瑞华潘为群潘为中
Owner HANGZHOU LONGYANG BIOTECH
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