Preparation method of pancreatin
A pancreatin and catalase technology, applied in pancreatin and other directions, can solve the problems of high cost of ultra-high pressure equipment, uncertain inactivation effect, and high production cost, and achieve the effect of enhancing virus safety.
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Embodiment 1
[0025] Embodiment 1, the preparation of trypsin comprises the following steps of carrying out successively:
[0026] (1) After slicing 100g of pancreas and 5g of duodenum, add 20g of anhydrous acetone, stir and mix, then grind with a colloid mill, and autolyse for 4 hours;
[0027] (2) Add 0.5 g of commercially available hydrogen peroxide solution hydrogen peroxide with a mass concentration of 30%, and stir at 2°C for 8 hours;
[0028] (3) Add 100000u unit catalase and stir at 5°C for 2h;
[0029] (4) Put the treatment solution treated in step (3) into a degreasing bucket, add pre-cooled raw acetone with a temperature of 0~5°C and a specific gravity ≤0.800, degrease for 2 hours, and open the valve to release degreasing acetone;
[0030] (5) Add calcium chloride as an activator to the treatment solution after step (4) for activation, stir evenly, put it into an extraction tank, and activate it for 20 hours at a temperature of 5°C;
[0031] (6) Separate and filter the activate...
Embodiment 2
[0037] It is basically the same as Example 1, except that 8 g of commercially available hydrogen peroxide solution with a mass concentration of 30% is added, and the amount of catalase added is 500,000 u. After testing, the prepared trypsin powder contained 1500u / g trypsin and 10000u / g pancrelipase, and no catalase residue.
[0038] Detection method of virus inactivation effect
[0039] After step (1) and before step (2), add 6.5 log 10 TCID50 / 0.1ml titer of the indicator virus EMCV (encephalomyocarditis virus, the amount of virus solution added is 5% of the volume of the solution, about 5ml of virus solution is added to 100g of pancreas).
[0040] The samples before adding hydrogen peroxide and after adding catalase reaction were taken out and inoculated into Vero cell culture respectively to observe the cytopathic effect (CPE). The effect of the sample before adding hydrogen peroxide on the cytopathic Figure 4 , the effect of the sample after adding catalase reaction on c...
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