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Method for separating heparitin from heparin by-product waste protein

A technology for heparin by-products and proteins, which is applied in the field of separating heparinoids from heparin by-products, can solve the problems of difficult separation of effective substances, waste of resources, etc., and achieves the effects of low cost, cost saving, and solving financial and material resources.

Active Publication Date: 2015-11-11
ZHEJIANG CASING ANIMAL BY PROD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the difficulty in separating the effective substances, these by-products are generally discarded or stored after the production of heparin, resulting in waste of resources

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Take 8L of heparin by-product waste protein and dissolve it with 16L of water. According to the ratio of adding 6g of resin per liter of protein, add 48g of resin for adsorption. The adsorption temperature is 59°C, the pH is 7.67, the salt concentration is 0.75mol / L, and the adsorption time is 8h. Collect the resin after adsorption, add 0.82mol / L sodium chloride solution for washing, add 4.5mol / L sodium chloride solution for elution after washing for 20 minutes, collect eluent after 4 hours of elution, add 300ml 90% ethanol, precipitation 16h. After the precipitation, the ethanol in the upper layer was poured off, and 180ml of water was added for dissolution. Use 30% sodium hydroxide to adjust the pH to 11.22, add 2ml of 30% hydrogen peroxide, stir evenly, and the oxidation time is 6 hours. After the oxidation is completed, use 8% hydrogen chloride to adjust the pH to 6.05, and add 40% hydrogen sulfite drop by drop Sodium, use Merck test paper to judge whether there a...

Embodiment 2

[0027] Take 8L of heparin by-product waste protein and dissolve it with 16L of water. According to the ratio of adding 8g of resin per liter of protein, 64g of resin was added for adsorption. The adsorption temperature was 59°C, the pH was 7.79, the salt concentration was 0.78mol / L, and the adsorption time was 8h. Collect the resin after adsorption, add 0.82mol / L sodium chloride solution for washing, add 4.5mol / L sodium chloride solution for elution after washing for 20 minutes, collect eluent after 3 hours of elution, add 300ml 90% ethanol, precipitation 16h. After the precipitation, the ethanol in the upper layer was poured off, and 180ml of water was added for dissolution. Use 30% sodium hydroxide to adjust the pH to 11.3, add 2ml of 30% hydrogen peroxide, stir evenly, and the oxidation time is 6 hours. After the oxidation is completed, use 8% hydrogen chloride to adjust the pH to 6.83, and add 40% hydrogen sulfite drop by drop Sodium, use Merck test paper to judge whethe...

Embodiment 3

[0031] Take 10L of heparin by-product waste protein and dissolve it in 25L of water. According to the ratio of adding 7g of resin per liter of protein, 70g of resin was added for adsorption. The adsorption temperature was 59°C, the pH was 7.96, the salt concentration was 0.78mol / L, and the adsorption time was 6h. Collect the resin after adsorption, add 0.82mol / L sodium chloride solution for washing, add 4.5mol / L sodium chloride solution for elution after washing for 20 minutes, collect eluent after 4 hours of elution, add 410ml 90% ethanol, precipitation 16h. After the precipitation, the ethanol in the upper layer was poured off, and 360ml of water was added for dissolution. Use 30% sodium hydroxide to adjust the pH to 11.36, add 4ml of 30% hydrogen peroxide, stir evenly, and the oxidation time is 9 hours. After the oxidation is completed, use 8% hydrogen chloride to adjust the pH to 7, and add 40% hydrogen sulfite drop by drop Sodium, use Merck test paper to judge whether t...

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Abstract

The invention relates to a method for separating heparitin from heparin by-product waste protein. The objective of the invention is to provide the method for separating heparitin from heparin by-product waste protein so as to realize reutilization of the waste and conservation of cost; and the method has the characteristics of simple process and low cost. According to a technical scheme in the invention, the method is characterized by comprising the following steps: 1) preparing a solution from the heparin by-product waste protein; 2) adding an adsorbent into the solution for adsorption; 3) washing and eluting the adsorbent, collecting eluate and carrying out precipitation so as to obtain a solid precipitate 1; 4) preparing a solution from the solid precipitate 1 and successively carrying out pH value adjustment, oxidation, pH value adjustment, reduction, pH value adjustment, salinity adjustment and precipitation so as to obtain a solid precipitate 2; and 5) drying the solid precipitate 2 so as to obtain heparitin.

Description

technical field [0001] The invention relates to a preparation method of heparin, in particular to a method for separating heparin from heparin by-products. Background technique [0002] Heparinoids refer to natural, synthetic and semi-synthetic substances that have similar effects to heparin. Heparinoids can combine with hundreds of proteins, affecting the spatial conformation, lifespan and biological activity of proteins. Heparinoids can also interact with extracellular Various proteins in the matrix combine to control their distribution, migration and information transmission. It can be seen that heparinoid is an important information molecule, which regulates the behavior of host cells, such as cell proliferation, differentiation, and combination, and participates in the identification and control of foreign cells. Heparin-like products can be used as raw materials for the production of sulodexide, Guanxinshu and other drugs, which are widely recognized by patients becau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/10
Inventor 高秉学童永刚潘卫春汤彬彩杨志秋江玲超
Owner ZHEJIANG CASING ANIMAL BY PROD CO LTD