Heparin sodium in living organisms and proteins can bind to live together, using salts to make it and the protein structure apart, called the process of halolysis.
HEPARINS Manufacturer describes the following steps:
1) pretreatment of D204 resin:
The resin is placed in a larger container, washed with clean and pure water, drained, and soaked in 80-90% industrial alcohol for about 24 hours, with the aim of washing the organic impurities from the resin and draining.
Rinse with 40-50℃ hot water repeatedly until alcohol free, and then drain.
Soak 2 times the resin in 4% hydrochloric acid solution for 2-3 hours, rotate constantly, and rinse with pure water or tap water until it is nearly neutral.
2) Preparation of hydrolysate:
Take fresh intestinal mucosa, add 5.5% sodium chloride, put into the pot, mix, adjust pH to 9.0 with industrial liquid alkali, keep stable for 10-15 minutes. Method 1: direct heat, raise the temperature to 100℃, keep the heat for 10 minutes, and discharge the material while it is hot; Method two: heat up to 50℃-55℃ within 30 minutes and hold for 2 hours. After 40 minutes, raise the temperature to 95℃, keep warm for 10 minutes, and remove the material while it is hot. Filtered with a nylon mesh with an aperture of 100-120 mesh, the residue was removed, and the filtrate was collected for hydrolysis.
3) Ion exchange:
The new D204S resin or the pretreated chlorine-type D204 resin was put into the hydrolysate at 50-60℃ (pH was controlled at 8.5-9.0, and the pomme degree of sodium chloride was controlled at about 4.8-5.2), and the mixed adsorption was carried out at this temperature for 6 hours. At this point, the concentration of residual solution after adsorption is less than 4U/ mL.
4) wash
The 7% NaCl solution with the same component as the resin can be washed and mixed with the resin twice. Each time we kept away for one and a half hours, the washing solution was poured to remove the low molecular weight heparin in the resin and impurities on the surface of the adsorbent material.
5) elution
With 0.8 times the weight of the resin 20% sodium chloride solution, if conditions permit, can be heated to 50℃, the first elution 6~8 hours, with 22-25% sodium chloride solution second elution 6 hours, the second eluent can be used for the next elution, eluent through filtration to remove fine particle impurity precipitation.
6) deposit
Add 90-95% alcohol to the eluent, control the alcohol content to 35-40 ° C (note that the salinity of the eluent is < 15 ° C), mix immediately, close the lid and seal, leave to settle for 24 hours, then siphon out the supernatant (the alcohol in the supernatant can be recovered), take the bottom precipitate, namely the crude heparin, and place it in a clean container.
7) dehydration
An anhydrous or ethanol was added to the drained and mashed heparin for dehydration, which was repeated for two to three times and then placed in a drying oven at 60-70℃. During the process, heparin should be frequently turned over to avoid agglomeration and moisture return. The crude heparin was obtained for analysis.
8) after heavy
1 kg of crude heparin +20 kg of water +3 kg of 22-25% NaCl solution, dissolve, heat, boil, filter, and add an alcohol precipitation, the alcohol level is generally controlled at 40 degrees is appropriate, the combination of heparin deposition is China's fine application of heparin, dehydration, deposition, drying and so on.
9) Rinse with D204S resin with clean water for several times, drain, soak in saturated brine for 24 hours, collect brine (including heparin), soak in hydroresin, prepare for reuse.