Is the project of extracting heparin sodium from pig small intestine feasible?

Heparin sodium is an important biochemical drug, which is used in clinical treatment of acute myocardial infarction and viral hepatitis. Combined with ribonucleic acid can increase the curative effect on hepatitis B. Combined with chemotherapy, it is helpful to prevent cancer cell metastasis. In addition, it also has the effect of preventing thrombosis. The production of heparin sodium is in short supply and there is a great gap in the international market. The intestinal mucosa of about 2000 pig small intestines after processing casings can produce 1 kg of heparin sodium, worth about 22000 yuan. Therefore, the use of pig intestinal mucosa as a by-product to produce heparin sodium has considerable value, obvious economic and social benefits, and broad prospects.

The specific process is as follows: production process

1. Extraction. Add 5% sodium chloride solution to the mucous membrane scraped off the pig casings to completely submerge the mucosa. The pH value of the solution was adjusted to 9.0-9.5 with sodium hydroxide solution and impregnated for about 10 hours. The key to this step is that the intestinal mucous fluid must be fresh and moderate in water content (the total solid content is 6%-7%).

2. Adsorption. Cool the above solution to about 50 ℃ to remove the oil and impurities floating on it. The treated 714 resin was added, and the amount of resin was 4%-6% of the mucous membrane. Stirring adsorption for 8 hours to 10 hours, adjust the PH value at any time to make it between 9.0 and 9.5. Then filter with 70 ~ 80 mesh nylon mesh bag to filter out the resin and remove the filtrate.

3. Washing. Rinse the resin adsorbed heparin sodium with water (until the lotion is clarified) and filter dry. Then put the resin in a bucket, add 1 times the amount of 1.4 mole / liter sodium chloride solution, stir and wash for 1 hour, filter dry. Then wash the resin twice with 1-fold 1.2 mole / liter sodium chloride solution and filter the resin dry. It is best to use refined salt water in the last washing.

4. Elution. The resin was stirred and eluted with 5 mole / liter sodium chloride for 2 hours, and the amount of sodium chloride was half of the amount of resin. Then filter and collect the filtrate. The resin was stirred and washed twice with 3 mole / liter sodium chloride solution for 24 hours each time (the amount of sodium chloride solution was still half of the amount of resin), filtered and collected.

5. Precipitation. Merge the filtrate for three times, adjust the PH value to 7.2-7.4, add 95% ethanol solution and stir evenly, and rest and precipitate. The amount of ethanol used varies from season to season. The ratio of filtrate to ethanol is 1:1 in winter, 1.3in spring, and 1.5in summer and autumn. The precipitation time is more than 24 hours, but not more than 48 hours, when there are obvious white particles precipitated and separated.

6. Dehydration and drying. The ethanol solution floating on the upper layer of heparin sodium precipitation was removed by siphon method. The precipitation was repeatedly washed with 95% ethanol for several times, collected, filtered and dried in vacuum, and then repeatedly dehydrated with acetone for 2 ~ 3 times. The ratio of acetone dosage to heparin sodium was 1:3. After drying, crude heparin sodium was obtained by drying at 60 ℃ or vacuum drying for more than 48 hours.

7. Purification: dissolve the crude product with 2% sodium chloride solution and prepare about 10% heparin sodium solution. Then adjust the PH value to 8, add 4% potassium permanganate or 30% hydrogen peroxide to decolorize, and remove alkaline and acidic proteins. Filter to adjust the PH to 6.5, then precipitate heparin sodium with 0.9 times 95% ethanol, filter out the precipitation, and drain until dry. Then dehydrated with anhydrous ethanol and acetone, finally treated with ether, and then dried in vacuum to obtain heparin sodium.