METHOD OF OBTAINING HEPARIN Russian patent published in 2023 - IPC C12P19/28 C08B37/10 A61P7/02 

FIELD: proteoglycan biotechnology.

SUBSTANCE: invention relates to a method of the synthesis of the anticoagulant heparin. The method of producing heparin is divided into two stages. At the first stage, the mucosa is diluted with water in a container for protein digestion in a ratio of 1:4, the resulting solution is heated to 25°C and add sodium hydroxide and sodium chloride to pH 9.0 and 3.2° according to Boma (°Be), the resulting solution is heated to 55°C, NaOH and NaCl are re-added until the solution pH is 9.0 and 3.2°Be, the 0.01% of protease by weight of the raw material is added, left in the mode of maintaining temperature equal to 55°C and continuous stirring for 1.5 hours, then heated to 95°C for 30 minutes, left in the mode of maintaining temperature equal to 95°C for 15 minutes, then the solution is cooled to a temperature of 58°C, after cooling, the solution is passed through a three-phase separator, pumped into a container for adsorption of sodium heparin ion, then AMBERLITE FPA98C1 resin is added to the solution in an amount of 3 wt.% of the raw material and stirred for 10 hours, the solution is poured through an 80 mesh, resin is collected and washed with running clean water until pH 7.0, then placed in a container to isolate the sodium heparin ion, the resin is mixed with a solution of sodium chloride with a density of 24°Be in a volume ratio of 1:1 to a salt concentration in solution of at least 20°Be, the resulting solution is heated to 50°C, then sodium chloride is added again until the salt concentration of the solution becomes at least 20°Be in relation to the volume of AMBERLITE FPA98Cl resin 1:1, left in the mode of maintaining temperature equal to 50°C for 4 hours, stirring continuously during the period of heating and maintaining temperature, then the liquid is separated from the resin from the solution using an 80 mesh screen. The resulting liquid is mixed with alcohol of at least 90% strength in a 1:1 ratio and left for 24 hours, resulting in a precipitate of raw heparin, which is filtered off and the filtrate is collected in a container. At the second stage, the solution obtained during the first stage is filtered using a microfilter with particle retention of 0.5 μm at a pressure of 0.25 MPa, after which the solution is filtered again using an ultrafiltration membrane with particle retention of 30,000 Da at a pressure of 1.0 MPa. The resulting solution is filtered through a nanofilter with particle retention of 3,000 Da at a pressure of 1.5 MPa, the unfiltered solution is collected using 6 M hydrochloric acid, the pH of the solution is adjusted to 7.0, then 95% alcohol is added in a ratio of 1:3, mixed for 5 minutes, allowed to stand for 12 hours, after which the precipitate is removed onto a freeze dryer and, after drying, dry heparin is obtained with a specific activity of at least 180 IU/mg.

EFFECT: ensuring continuity of the technological process for producing of heparin.

2 cl