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High-extraction rate preparation method of high-activity hepatocyte growth-promoting factor

A technology of hepatocyte growth factor and extraction rate, applied in the field of preparation of high extraction rate and high activity hepatocyte growth-promoting factor, can solve the problems of loss of active substance content, low extraction rate, low activity of extract, etc., and achieve high yield , the effect of improving filtration efficiency

Inactive Publication Date: 2015-03-04
HEFEI PINGGUANG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The process of this preparation method is cumbersome, and the eluent with salt needs to be eluted and then desalted. These operating steps will lose the content of active substances, resulting in low activity of the extract and low extraction rate.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The preparation method of a high extraction rate and high activity hepatocyte growth-stimulating hormone proposed by the present invention comprises the following steps:

[0032]S1. Pretreatment of suckling pig liver: take suckling pig liver, remove gallbladder, fascia and connective tissue, wash with water, drain and then wash with water for injection, then cut into small pieces or thin strips with a width of 3-5cm, and then Put it in a meat grinder and grind it into suckling pig liver particles with a particle size of 1-3mm;

[0033] S2. Homogenization: Add suckling pig liver granules obtained in S1 into a colloid mill and grind at a speed of 3100rpm for 3.5 minutes to obtain suckling pig liver mince, and add suckling pig liver mince and water for injection into a mixer at a weight ratio of 5:5.5, and The rotating speed of 310rpm was stirred for 17min to obtain suckling pig liver homogenate;

[0034] S3, preparation of pepsin: take pig gastric mucosa and grind it int...

Embodiment 2

[0041] The preparation method of a high extraction rate and high activity hepatocyte growth-stimulating hormone proposed by the present invention comprises the following steps:

[0042] S1. Pretreatment of suckling pig liver: take suckling pig liver, remove gallbladder, fascia and connective tissue, wash with water, drain and then wash with water for injection, then cut into small pieces or thin strips with a width of 3-5cm, and then Put it in a meat grinder and grind it into suckling pig liver particles with a particle size of 1-3mm;

[0043] S2. Homogenization: add the suckling pig liver granules obtained in S1 to the colloid mill and grind at a speed of 3250rpm for 2min to obtain suckling pig liver mince. Stir at a rotating speed of 14min to obtain suckling pig liver homogenate;

[0044] S3. Preparation of pepsin: take the pig gastric mucosa and grind it into pig gastric mucosa and put it in a reaction container, add 1.5wt% hydrochloric acid solution, the volume ratio (g / m...

Embodiment 3

[0051] The preparation method of a high extraction rate and high activity hepatocyte growth-stimulating hormone proposed by the present invention comprises the following steps:

[0052] S1. Pretreatment of suckling pig liver: take suckling pig liver, remove gallbladder, fascia and connective tissue, wash with water, drain and then wash with water for injection, then cut into small pieces or thin strips with a width of 3-5cm, and then Put it in a meat grinder and grind it into suckling pig liver particles with a particle size of 1-3mm;

[0053] S2. Homogenization: Add suckling pig liver granules obtained in S1 into a colloid mill and grind at a speed of 3150rpm for 2.5 minutes to obtain suckling pig liver mince, and add suckling pig liver mince and water for injection into a blender at a weight ratio of 1.1:1, and The rotating speed of 320rpm was stirred for 16min to obtain suckling pig liver homogenate;

[0054] S3, preparation of pepsin: take the pig gastric mucosa and grind...

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Abstract

The invention discloses a high-extraction rate preparation method of a high-activity hepatocyte growth-promoting factor. The preparation method comprises the following steps: 1) performing pretreatment on a suckling pig liver; 2) homogenizing; 3) preparing pepsin; 4) performing enzymolysis: putting a homogenate of the suckling pig liver into a reaction container, dropping a hydrochloric acid solution till the pH value is 2.5-3.5, further adding the pepsin, stirring and then putting the reaction container and contents thereof into a water bath pot for hydrolysis to obtain an enzymolyzed material; 5) inactivating; 6) centrifuging; 7) performing coarse filtration: sequentially filtrating neutral centrifugal liquid with a hollow fiber ultrafiltration device with molecular weight cutoff of 100KD and a hollow fiber ultrafiltration device with the molecular weight cutoff of 10KD to obtain coarse filtrate; and 8) performing ultrafiltration: performing ultrafiltration on the coarse filtrate with an ultrafiltration membrane with the molecular weight cutoff of 6000D to obtain a hepatocyte growth-promoting factor solution. The preparation method disclosed by the invention has the advantages of simple and mild process, energy conservation, environment friendliness and high extraction rate; and the prepared hepatocyte growth-promoting factor is relatively high in activity and high in safety.

Description

technical field [0001] The invention relates to the technical field of hepatocyte growth-stimulating hormone, in particular to a preparation method of hepatocyte growth-stimulating hormone with high extraction rate and high activity. Background technique [0002] Hepatocyte growth-stimulating hormone is a small molecular weight polypeptide mixture extracted from fresh suckling pig liver, with a molecular weight below 10,000. Its pharmacological effects are mainly to stimulate the DNA synthesis of normal liver cells, promote the regeneration of liver cells, protect the damaged liver cells, promote the recovery of liver function, and also regulate the body's immune function and anti-hepatic fibrosis. It is clinically used to treat severe hepatitis, chronic active hepatitis and liver cirrhosis. At present, there have been many patents and literature reports on hepatocyte growth-stimulating hormone, a biological agent drug. [0003] In the patent ZL03116052.2, a hepatocyte gro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34
Inventor 王康林马婕陈玉海
Owner HEFEI PINGGUANG PHARMA
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