Extended introduction:
Production technology of heparin sodium
1. Raw material treatment: Clean fresh pig lung (or frozen pig lung after natural thawing) carefully with clear water to remove internal and external dirt and skin fat, then mince it into paste, fully stir it, add the same amount of water to mix, and then add a little preservative with solubility of .01%to mix evenly.
2. Enzymatic extraction: firstly, finely adjust the PH value of the above raw materials to 8-9 with a small amount of dilute alkali solution under full stirring (which can be measured by corresponding precise PH test paper, the same below), then add pre-ground fresh pancreatic pulp as enzymolysis agent (the appropriate amount of pig pancreatic pulp added is 1%- 1.5% of the actual weight of the raw material solution), and slowly heat up to 40 after stirring. Keep the PH of the feed solution at 7.5-8, keep the temperature at 37-40 degrees, perform enzymolysis for 3-4 hours, then raise the temperature to 47-50 degrees, keep the PH value at 8.0-8.5, add a small amount of porcine pancreatic pulp appropriately, and continue enzymolysis for 4-5 hours. In the above-mentioned enzymatic hydrolysis process, if the pH paper of the enzymatic hydrolysis solution is rechecked and decreased, a sentence should be given in time. Adjust the pH value of hydrochloric acid to 5.5-6, and then raise the temperature to 80 degrees. Adding refined salt (containing NaCL≥95%, calcium magnesium calcium salt)
Third, ion exchange adsorption; Firstly, the enzymolysis extract was cooled to room temperature, and a thin layer of oil floating on the liquid surface was carefully removed. The temperature was controlled to 45 degrees, and the heating was stopped. The heparin sodium component was added to the pretreated D-254 resin with stirring. The dosage of new resin is generally 2.55-3% of the feed liquid. After regeneration, it should be increased as appropriate, stirred and adsorbed for 3 hours, and the filtrate can be used for production and treatment of coronary heart disease. Rinse the D-254 resin containing heparin with clear water, and then use 1 times/0.2 mol/L sodium chloride solution to perform heparin sodium operation on the washed D-254 resin: the first eluent is about10.5 times the resin volume, and the second eluent is about 0.5 times the resin volume, with elution time. The resin was filtered to dryness, and the eluents were combined. Adjust the obtained eluent to PH= 10- 1 1, stir for 30 minutes, then let it stand for 6 hours, then carefully separate out the supernatant, and filter the lower sediment as dry as possible. Combine the supernatant and filtrate, finely adjust the PH to 6.0-6.5, add 95% ethanol with a ratio of 1.5 to precipitate overnight, carefully dry the supernatant the next day, collect the lower precipitate, and filter to dryness (mother liquor can be used as eluent before pH adjustment). Vacuum drying to obtain crude heparin sodium.
Fourthly, refining: the crude heparin sodium is completely dissolved in 2% sodium chloride solution to prepare a solution with a solubility of about 8%, and the temperature is appropriately increased in this process to facilitate dissolution. At night, finely adjust the pH value of the above substances to 8.0-8.2 with 5mol/L sodium hydroxide solution, raise the temperature to 78-80 degrees, and add 0. 1 .5-0.2 mol/L potassium permanganate solution per1100 million unit until the purple color no longer fades, which is the end point of the first oxidation operation; When the red color has just faded, add a little saturated sodium sulfite solution. After the above feed liquid is properly cooled, it is finely filtered once, and then filtered again when the temperature of the filtrate is reduced to 36 degrees. The pH of the filtrate is adjusted to 10.5- 1 1.0 with a small amount of saturated sodium hydroxide solution, and a small amount of 3%-5% hydrogen peroxide solution is slowly added under full stirring, and the temperature is 25-27℃. After the oxidation process, the feed liquid is filtered by a "degerming filter", the filtrate is adjusted to PH=5.8-6.5 with a little hydrochloric acid, and 95% ethanol with 0.9 times is added to precipitate at 5- 10 degree for 24 hours. Collect the above precipitate, dissolve it with a small amount of 10% sodium chloride solution, add 3-4 times of 95% ethanol for precipitation, and collect the precipitate (ethanol can be recycled after recovery, distillation and dehydration). Dehydrate that precipitate with absolute ethanol, grinding, dehydrate with acetone, grinding, dehydrating with acetone, and vacuum drying with far infrared ray (50-60 DEG C) to obtain refined heparin sodium.