Method for preparing deoxyribonucleic acid from waste liquid after heparin extraction of pig small intestinal mucosa

A technology of deoxyribose nucleic acid and mucosal heparin, applied in the field of preparation of deoxyribonucleic acid, can solve the problems of environmental pollution, waste of active substances in medical care, etc., and achieve the effect of simple preparation and process, rich raw materials and low cost

Active Publication Date: 2017-11-28
OCEAN UNIV OF CHINA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the intestinal mucosa extracts heparin, a large amount of waste is produced. The discharge of waste not only causes environmental pollution, but also wastes the active substances in the waste that have medical and health care functions.

Method used

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  • Method for preparing deoxyribonucleic acid from waste liquid after heparin extraction of pig small intestinal mucosa
  • Method for preparing deoxyribonucleic acid from waste liquid after heparin extraction of pig small intestinal mucosa
  • Method for preparing deoxyribonucleic acid from waste liquid after heparin extraction of pig small intestinal mucosa

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preparation example Construction

[0030] In a specific embodiment, the preparation method of deoxyribonucleic acid provided by the present invention is to perform column adsorption on the waste liquid after heparin extraction of porcine small intestinal mucosa, and then perform desorption, collect the desorption liquid to adjust pH, alcohol precipitation, dehydration, and drying, specifically The process steps are as follows:

[0031] (1) Column adsorption: The ion exchange resin is loaded into the adsorption column, and the waste liquid after the heparin extraction of the porcine small intestine mucosa is extracted by the pump is adsorbed through the adsorption column;

[0032] (2) Desorption: wash the adsorption column with sodium chloride with a concentration of 0.05 to 0.2 mol / L, after draining, elute with a sodium chloride solution with a temperature of 60 to 70 °C and a concentration of 2.0 to 4.0 mol / L, Collect the desorption solution;

[0033] (3) Acid precipitation: adjust the pH of the collected des...

Embodiment 1

[0040] Put 200g of Rohm and Haas AMBERLITE FPA 98 CL resin into the adsorption column, and use a pump to absorb 170kg of waste liquid after heparin extraction from the porcine small intestinal mucosa through the adsorption column, and first use a sodium chloride aqueous solution with a concentration of 0.1moL / L Wash 2 times the column volume, then elute 2 times the column volume with an aqueous solution of sodium chloride at a temperature of 60°C and a concentration of 4.0 mol / L, and collect the desorption solution. Adjust the pH to 2 with 3mol / L hydrochloric acid while stirring the collected desorption solution, and let it stand at 4°C for 4h. Centrifuge, remove the supernatant, wash the precipitate three times with distilled water, remove the supernatant residue, slowly add 2mo / L sodium hydroxide aqueous solution to the precipitate until the precipitate is completely dissolved, add 3 times of ethanol with a volume fraction of 90% for precipitation , standing at 4°C for 12 ho...

Embodiment 2

[0042] Put 500g of Rohm and Haas AMBERLITE FPA 98 CL resin into the adsorption column, and use a pump to extract 570kg of waste liquid from the porcine small intestinal mucosa after heparin is absorbed through the resin. After washing the resin, elute with 2.0 mol / L aqueous sodium chloride solution at 60°C for 2 times the column volume, and collect the desorption solution. Adjust the pH to 3 with 2mol / L hydrochloric acid while stirring the collected desorption solution, and let it stand at 4°C for 6h. Centrifuge, remove the supernatant, wash the precipitate three times repeatedly with distilled water, remove the supernatant residue, slowly add 2mo / L sodium hydroxide aqueous solution to the precipitate until the precipitate is completely redissolved, add 4 times the volume fraction of 95% ethanol Precipitate, stand at 4°C for 24 hours, centrifuge to remove the supernatant, and collect the lower layer of precipitate. Dissolve the precipitate in distilled water at 50°C, stir to ...

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Abstract

The invention relates to the field of preparation of DNA (desoxyribonucleic acid) from animal tissue, in particular to a method for preparing DNA from a waste liquid of porcine small intestinal mucosa after heparin extraction. According to the method, the waste liquid of the porcine intestinal mucosa after the heparin extraction is subjected to column adsorption, desorption, acid settlement, re-dissolution, alcohol precipitation and dialysis to obtain high-purity DNA, and the molecular weight of DNA is in a range from 3 kDa to 70 kDa. The waste liquid of the porcine small intestinal mucosa after the heparin extraction is selected as a raw material for extraction of DNA, the comprehensive utilization rate of the porcine small intestinal mucosa can be increased, the waste liquid emission is reduced, co-production of heparin and nucleic acid products in the porcine small intestinal mucosa is realized, the additional value of the raw material is increased, and the stable industrial raw material can be provide for DNA. The product obtained with the method has great market application value in the aspects of immune improvement, resistance to tumors, resistance to viruses, resistance to cardiovascular and cerebrovascular diseases and the like.

Description

technical field [0001] The invention relates to the field of preparing deoxyribonucleic acid from animal tissues, in particular to a method for preparing deoxyribonucleic acid from waste liquid extracted from pig small intestinal mucosa heparin. Background technique [0002] Nucleic acid is one of the most basic substances of life. It widely exists in all animals, plant cells, and microorganisms. Nucleic acid is often combined with protein to form nucleoprotein in organisms. Different nucleic acids have different chemical compositions and nucleotide arrangement sequences. According to different chemical composition, nucleic acid can be divided into ribonucleic acid (referred to as RNA) and deoxyribonucleic acid (abbreviated as DNA). [0003] DNA is the main substance for storing, replicating and transmitting genetic information, and plays an important role in the individual growth, development, reproduction, inheritance and other links of organisms, and can promote cell gro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 于广利孙学超陈璇刘亚男许春龙
Owner OCEAN UNIV OF CHINA
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