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.

Active Publication Date: 2018-11-13
苏州良辰生物医药科技有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

The disadvantage of this method is that the inactivation effect of high pressure treatment on some viruses is uncertain, and the ultra-high pressure equipment is expensive and the production cost is high, which is basically not advisable in production practice

Method used

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  • Preparation method of pancreatin
  • Preparation method of pancreatin
  • Preparation method of pancreatin

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to a preparation method of pancreatin. The preparation method comprises following steps: pancreas is subjected to grounding, hydrogen peroxide is added so as to obtain a mixture,wherein the mass ratio of hydrogen peroxide in the mixture is controlled to be 0.1 to 2%; the mixture is stirred for 1 to 8h, catalase is added for 0.5 to 2h of reaction, and degreasing, activating,deposition, and drying are carried out so as to obtain pancreatin. In the preparation method, hydrogen peroxide is capable of realizing full contact with virus in a relatively short time, so that virus inactivation is realized; the residual hydrogen peroxide is degraded by catalase into water and oxygen, so that no hydrogen peroxide is left in pancreatin, the drug virus safety is improved greatly,and the obtained pancreatin product is capable of satisfying the strict requirements of pharmacopeia and animal-sourced biochemical medicine standards of different states.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of trypsin. Background technique [0002] Pancreatin is a mixed enzyme preparation extracted from the pancreas of mammals such as pigs, cattle, and sheep. The main components are trypsin, pancreatic amylase, and pancreatic lipase, as well as carboxypeptidase, chymotrypsin, and elastase. , kallikrein and ribonuclease. As early as 1938, pancreatic enzyme products have been used for cystic fibrosis and chronic pancreatitis, and most patients need to take it for life. At present, pancreatin is a digestive aid drug recorded in the pharmacopoeias of various countries, and it is clinically used to treat indigestion, loss of appetite, and digestive disorders caused by hepatopancreatic diseases. Since pancreatin contains various active substances, it can be used as raw material medicine to extract required biochemical substances therefrom. In addition, tryp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/94
CPCC12N9/94
Inventor 殷文静
Owner 苏州良辰生物医药科技有限公司
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