Heparin sodiumHow is heparin sodium extracted

Details:

Heparin sodium is a kind of mucopolysaccharide sulfate anticoagulant. It is the sodium salt of csso3 extracted from the intestinal mucosa of pigs or cattle. It belongs to mucopolysaccharide substance. It is the most effective anticoagulant in the world. This paper mainly introduces a kind of technology of extracting heparin sodium, using pig intestinal mucosa as raw material, through biological enzymatic hydrolysis technology, the method of extracting heparin sodium.

Heparin sodium

The extraction process of heparin sodium is as follows:

1, enzymatic hydrolysis: take pig's small intestinal mucous membrane as raw material into the agitator, add the raw material weight 0.5% amylase (35 thousand U/g), mix and then adjust to pH 8.0-9.0, warm up to 45-50 degrees Celsius, after constant temperature stirring enzymolysis for 2.8 hours, we need to control the pH value of the reaction system stable during enzymatic hydrolysis.

2. Enzymolysis: the hydrolysate was heated to 90 ℃ for 10 min, then cooled immediately.

3. Filtration: the quench liquid is filtered twice with a filter. First, the filter membrane with a pore size of 2 μ m is used for coarse filtration, and then the membrane with a pore size of 0.22 μ m is used for filtration to get the filtrate after removing impurities.

4. Enzymolysis protein material: adjust the filtrate to pH 7.5-9.0, then raise the temperature to 45-55 ℃, add 0.5% flavor protease (38000 U / g) by weight of raw material, stir the enzymolysis at 45-55 ℃ for 4-6 hours, control the pH value of the reaction system to be stable, use the enzymolysis solution to raise the temperature to 90 ℃, keep it for 10 minutes in the process of enzymolysis inactivation, and then immediately cool to obtain quenching liquid.

5. Remove protein: remove protein with a 5 micron filter and concentrate the quenching solution.

6. Deposition: 95 wt% ethanol was added in the liquid concentrator with equal density, and the supernatant was removed after 10 hours of stirring. Anhydrous ethanol was added in the deposition to dehydrate and freeze-dried to obtain crude heparin sodium.

7. Refining: adjust the pH of pure water added 9 times the weight of crude heparin sodium to 1.5, then dissolve the crude product, filter and remove impurities, adjust the pH of the filtrate to 11, record the volume of the solution at this time, add 30wt% hydrogen peroxide according to 4% of the volume, maintain oxidation at 25 ℃, and pay attention to control the pH of the reaction system to be stable at 11. After 30 hours of oxidation, the pH was adjusted to 6.5. At the same time, 95wt% ethanol was added to the solution for 10 hours after stirring. The supernatant was removed and the ISO density absolute ethanol was dehydrated in the precipitation. The ethanol precipitates were filtered out, and the residual ethanol was evaporated, evaporated and precipitated.

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