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Novel extraction method of cerebroprotein hydrolysate solution

A technology of brain protein hydrolyzate and extraction method, which is applied in the fields of hydrolyzed protein components, drug combinations, nervous system diseases, etc., and can solve problems such as tryptophan content not meeting quality standards, incomplete hydrolysis, and identification items not meeting regulations, etc.

Active Publication Date: 2012-01-04
HANGZHOU HUADI GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest disadvantage of this method of hydrolysis is that the hydrolysis is not complete, which directly affects the amino acid content of the product, so additional amino acids need to be added in the subsequent production process to meet the national standards.
[0005] 2. Enzyme method + acid hydrolysis extraction method: This method is based on the multi-enzyme extraction method, and continues to use acid hydrolysis, generally using 6M hydrochloric acid to hydrolyze, so as to achieve the purpose of increasing the amino acid content; but the biggest disadvantage of this method The reason is that acid hydrolysis will destroy the tryptophan in the product, so the content of tryptophan in the product quality standard does not meet the quality standard, and the identification items in the product quality standard also do not meet the requirements
[0006] 3. Enzyme method + alkali hydrolysis extraction method: This method is also based on the multi-enzyme extraction method, and then continues to use alkali hydrolysis, generally using 6M sodium hydroxide to hydrolyze, so as to achieve the purpose of increasing the amino acid content; but this method The biggest disadvantage is that although alkaline hydrolysis will not destroy the tryptophan in the product, the structure of the remaining amino acids in the product will be racemized due to alkaline hydrolysis, thus turning into D-type amino acids that are useless to the human body
Therefore, it has a great influence on the curative effect of the product

Method used

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  • Novel extraction method of cerebroprotein hydrolysate solution
  • Novel extraction method of cerebroprotein hydrolysate solution

Examples

Experimental program
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Effect test

Embodiment 1

[0025] A method for extracting cerebroprotein hydrolyzate solution, comprising the following steps:

[0026] ①. Take fresh or frozen porcine brain, remove the cerebellum and brainstem, and remove the fascia on the surface of the brain tissue, wash it with purified water, add 1 times the purified water colloid and grind for homogenization 3 times.

[0027] ② Take the homogenate, heat it to 45°C, adjust the pH value to 7.5 with sodium hydroxide, add 2.5% trypsin, and stir for 2 hours. After the reaction, the pH value was adjusted to 2.5 with hydrochloric acid, heated to 95° C., kept for 30 minutes, and the supernatant was collected by centrifugal filtration.

[0028] ③. Take the centrifuged supernatant, add 50wt% hydroxylamine solution in a volume ratio of 1:1, adjust the pH value to 2.5 with hydrochloric acid, and react in a closed manner at 80°C for 30 hours. After the reaction, cool to 30°C and adjust the pH value with sodium hydroxide To 7.5, centrifuge to get the supernata...

Embodiment 2

[0031] A method for extracting cerebroprotein hydrolyzate solution, comprising the following steps:

[0032] ①. Take fresh or frozen pig brain, remove the cerebellum and brainstem, and remove the fascia on the surface of the brain tissue, wash it with water for injection, add 2 times purified water or water for injection colloid mill to homogenize twice;

[0033] ② Take the homogenate, heat it to 35°C, adjust the pH value to 8.5, add 0.5% trypsin, stir and react for 8 hours; adjust the pH value to 2.5, heat to 94°C, keep warm for 10 minutes, and centrifuge;

[0034] ③. Take the centrifuged supernatant in step ②, add 40wt% hydroxylamine solution at a volume ratio of 3:1, adjust the pH value to 2.5, and react in a closed manner at 90°C for 20 hours; cool to 33°C, adjust the pH value to 7.2, and centrifugally filter;

[0035] ④, take step ③ centrifuged supernatant, ultrafiltration with ultrafiltration membrane with a molecular weight of 8KD, and collect the filtrate; according to...

Embodiment 3

[0037] A method for extracting cerebroprotein hydrolyzate solution, comprising the following steps:

[0038] ①. Take fresh or frozen pig brain, remove the cerebellum and brainstem, and remove the fascia on the surface of the brain tissue, wash it with water for injection, add 1.5 times of purified water or water for injection colloidal grinder for 3 times;

[0039] ② Take the homogenate, heat it to 40°C, adjust the pH value to 8.0, add 1% trypsin, stir and react for 5 hours; adjust the pH value to 2.2, heat to 98°C, keep warm for 20 minutes, and centrifuge;

[0040] ③. Take the centrifuged supernatant in step ②, add 45wt% hydroxylamine solution at a volume ratio of 1:3, adjust the pH value to 2.0, and react in a closed manner at 92°C for 15 hours; cool to 37°C, adjust the pH value to 6.9, and centrifugally filter;

[0041] ④. Take the centrifuged supernatant from step ③, ultrafilter with an ultrafiltration membrane with a molecular weight of 12KD, and collect the filtrate; acc...

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Abstract

The invention discloses an extraction method of a cerebroprotein hydrolysate solution. In the extraction method, the extraction of a cerebroprotein hydrolysate solution is realized by combining multi-enzyme hydrolysis and hydroxylamine cleavage; based on the multi-enzyme extraction method, the method subsequently utilizes hydroxylamine for cleavage, thereby increasing the amino acid content and simultaneously retaining tryptophan without changing the structures of amino acids; by utilizing the hydroxylamine, peptide bonds between asparagine and glycin can be specifically cleaved; under acidicconditions, peptide bonds between asparagine and proline can be cleaved; and peptide bonds between asparagine and leucine as well as peptide bonds between asparagine and alanine can be partially cleaved. The extraction method improves the total amino acid content and the peptide content in a cerebroprotein hydrolysate injection, thereby improving the quality standard of the product, and ensuring that the product meets the national drug standards.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting a cerebroprotein hydrolyzate solution. Background technique [0002] Cerebroprotein hydrolyzate injection stock solution is a light yellow clear liquid, which is obtained by extracting, separating and refining pig brain tissue. It contains about 16 kinds of free amino acids and a small amount of peptides; it is a kind of peptide energy neurotrophic unique to the brain Drugs can act on the central nervous system in various ways, regulate and improve the metabolism of neurons, promote the formation of synapses, induce the differentiation of neurons, and further protect nerve cells from various ischemia and neurotoxin damage. Brain protein hydrolyzate injection stock solution can pass the blood-brain barrier, promote the synthesis of proteins in the brain, affect the respiratory chain, have the ability to protect against hypoxia, improve energy metabolism in the ...

Claims

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Application Information

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IPC IPC(8): A61K38/01A61P25/28
Inventor 周正兵高留根刘保林
Owner HANGZHOU HUADI GRP CO LTD
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