Method for purifying dermatan sulfate from heparin byproduct

A technology of heparin by-product and dermatan sulfate, applied in the field of purifying dermatan sulfate from heparin by-product, can solve the problems such as complicated steps for extracting dermatan sulfate, unsuitable for industrial large-scale production, unable to meet market demands, and the like, and the cost is achieved. Low, easy to scale up, high product purity effect

Active Publication Date: 2010-11-17
SHENZHEN HEPALINK PHARMA GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Dermatan sulfate is extracted from animal tissues at the expense of animals, and the amount of dermatan sulfate obtained is relatively small, which canno...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Dissolve heparin by-products into a 20% solution, add potassium acetate to it to a concentration of 20%, stir to dissolve, adjust the pH to 5 with acetic acid, precipitate at 2°C for 1 day, and collect the supernatant by centrifugation to obtain a sulfated polysaccharide solution after removing heparin.

[0019] Add 0.5 times the volume of Benedict's reagent and 0.05 times the volume of saturated sodium hydroxide to the aforementioned sulfated polysaccharide solution, stir and precipitate at room temperature for 10 minutes, and centrifuge to separate the supernatant and the precipitate. Add 0.5 times the volume of Banner's reagent to the precipitate, stir well to dissolve, then add 0.05 times the volume of saturated sodium hydroxide, stir and precipitate at room temperature for 10 minutes, and centrifuge to collect the precipitate.

[0020] Dilute the obtained precipitate with 400ml of purified water, adjust the pH to be neutral, put on a DEAE-Sepharose FF chromatography...

Embodiment 2

[0022] Dissolve heparin by-products into a 10% solution, add potassium acetate to it to a concentration of 80%, stir to dissolve, adjust the pH to 7 with acetic acid, precipitate at 2°C for 7 days, and collect the supernatant by centrifugation to obtain a sulfated polysaccharide solution after removing heparin.

[0023] Add 1.6 times of Banner's reagent and 0.2 times of saturated sodium hydroxide to the aforementioned sulfated polysaccharide solution, stir and precipitate at room temperature for 60 minutes, and centrifuge to separate the supernatant and the precipitate. Add 1.6 times of Banner's reagent to the precipitate, stir thoroughly to dissolve, then add 0.2 times of saturated sodium hydroxide, stir and precipitate at room temperature for 60 minutes, and centrifuge to collect the precipitate.

[0024] Dilute the obtained precipitate with 400ml of purified water, adjust the pH to be neutral, put on a DEAE-Sepharose FF chromatography column, wash 300ml with 0.6M sodium chlo...

Embodiment 3

[0026] Dissolve heparin by-products into an 8% solution, add potassium acetate to it to a concentration of 60%, stir to dissolve, adjust the pH to 6.5 with acetic acid, precipitate at 5°C for 5 days, and collect the supernatant by centrifugation to obtain a sulfated polysaccharide solution after removing heparin.

[0027] Add 0.8 times of Banner's reagent and 0.1 times of saturated sodium hydroxide to the aforementioned sulfated polysaccharide solution, stir and precipitate at room temperature for 40 minutes, and centrifuge to separate the supernatant and the precipitate. Add 0.8 times of Banner's reagent to the precipitate, stir thoroughly to dissolve, then add 0.1 times of saturated sodium hydroxide, stir and precipitate at room temperature for 40 minutes, and centrifuge to collect the precipitate.

[0028] Dilute the obtained precipitate with 400ml of purified water, adjust the pH to be neutral, put on a DEAE-Sepharose FF column, wash 300ml with 0.3M sodium chloride, elute 4...

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Abstract

The invention discloses a method for purifying dermatan sulfate from a heparin byproduct. The method comprises the following steps of: using the heparin byproduct as a raw material; dissolving the heparin byproduct, and then adding potassium acetate into the dissolved heparin byproduct; adjusting the pH value of solution by using acetic acid to generate a deposit, wherein clear solution is sulfate polysaccharide solution; adding a Ban's agent and saturated sodium hydroxide solution into the sulfate polysaccharide solution; centrifugally collecting the deposit; performing re-precipitation; dissolving the deposit again; washing away copper ions by using a chromatography column with anion exchange properties and sodium chloride solution; performing linear gradient elution by using the sodium chloride solution; collecting eluant; and evaporating and concentrating the eluant, and depositing the eluant by using ethanol to obtain high-purity dermatan sulfate. The method has the advantages of simple purification process, high purification yield, low cost, easy amplification and suitability for industrial production.

Description

technical field [0001] The invention relates to a preparation method of dermatan sulfate, in particular to a method for purifying dermatan sulfate from heparin by-products. Background technique [0002] Dermatan sulfate (DS for short) is a natural glycosaminoglycan composed of repeating disaccharide units. The disaccharides are acetamidogalactose and iduronic acid, mainly N-acetyl A double sugar chain structure composed of galactosamine 4-sulfate and L-iduraldehyde. DS is widely distributed in animal tissues. It has the functions of antithrombotic, anti-edema, promoting skin renewal, preventing hyperkeratinization and acanthosis of the skin, moisturizing the skin, and maintaining moisture. It has been paid more and more attention by domestic and foreign experts in recent years. [0003] For a long time, animal cortex has been the main raw material for preparing dermatan sulfate, and it is also the main source of commercial dermatan sulfate. As early as the 1960s, Clifonell...

Claims

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Application Information

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IPC IPC(8): C08B37/00
Inventor 李坦马小来李锂
Owner SHENZHEN HEPALINK PHARMA GRP CO LTD
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