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A method for removing keratan sulfate from chondroitin sulfate crude extract

A technology of chondroitin sulfate and keratan sulfate, which is applied in the biological field, can solve problems such as difficult industrial production and removal of keratan sulfate, and achieve the effects of simple operation, solving the difficulty of removing keratan sulfate, and saving usage

Active Publication Date: 2018-08-07
RIZHAO LANSHAN BIOCHEM PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the problem that it is difficult to remove keratan sulfate from chondroitin sulfate and industrialized production, the present invention provides a method for removing keratan sulfate from chondroitin sulfate crude extract, which has good removal efficiency and the obtained chondroitin sulfate High purity, simple operation, suitable for industrial production

Method used

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  • A method for removing keratan sulfate from chondroitin sulfate crude extract

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Experimental program
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Effect test

Embodiment 1

[0021] Taking the chondroitin sulfate crude extract and adding deionized water to prepare 10L mass fraction is a solution of 10%, the conductivity of the solution is 20ms / cm, wherein the chondroitin sulfate content is 90% in the crude extract, and the content of keratan sulfate is 2%, the protein content is 1%,. Use 2moL / L sodium hydroxide solution to adjust the pH to 12, stir at room temperature for 2 hours, then adjust the pH to 8 with hydrochloric acid, raise the temperature to 45°C, keep it warm for 3 hours, lower to room temperature, adjust the pH of the solution to 6 with hydrochloric acid, and filter to obtain the filtrate. Add solid sodium chloride to the filtrate to adjust the conductivity of the solution to 45ms / cm, and add 95% ethanol solution very slowly to the solution with adjusted conductivity, control the flow rate to 15mL / (L·min), and continuously Stir until the alcohol content of the solution is 50%, let it stand for 2 hours, filter to obtain a precipitate, d...

Embodiment 2

[0023] Taking the chondroitin sulfate crude extract and adding deionized water to prepare 10L mass fraction is a solution of 15%, the conductivity of the solution is 22ms / cm, wherein the chondroitin sulfate content is 92% in the crude extract, and the content of keratan sulfate is 4%, and the protein content is 3%. Use 2moL / L sodium hydroxide solution to adjust the pH to 14, stir at room temperature for 3 hours, then adjust the pH to 9 with hydrochloric acid, raise the temperature to 50°C, and keep it warm for 4 hours, then lower the temperature to room temperature and adjust the pH of the solution to 7 with hydrochloric acid, and filter to obtain the filtrate. Add solid sodium chloride to the filtrate to adjust the conductivity of the solution to 48ms / cm, and add absolute ethanol solution to the solution with adjusted conductivity very slowly, control the flow rate to 10mL / (L min), and keep stirring, Until the alcohol content of the solution is 55%, let it stand for 4 hours, ...

Embodiment 3

[0025] Taking the chondroitin sulfate crude extract and adding deionized water to prepare 10L mass fraction is a solution of 15%, the conductivity of the solution is 21ms / cm, wherein the chondroitin sulfate content is 90% in the crude extract, and the content of keratan sulfate is 2%, protein content is 1%. Use 2mol / L sodium hydroxide solution to adjust the pH to 14, stir at room temperature for 3 hours, then adjust the pH to 9 with hydrochloric acid, raise the temperature to 50°C, and keep it warm for 4 hours, then lower to room temperature and adjust the pH of the solution to 6 with hydrochloric acid, filter, and then filter Add solid sodium chloride to the solution to adjust the conductivity of the solution to 46ms / cm, add absolute ethanol solution to the solution with adjusted conductivity very slowly, control the flow rate to 13mL / (L·min), and keep stirring until The alcohol content of the solution is 55%, let it stand for 4 hours, and filter to obtain the precipitate, wh...

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Abstract

The invention belongs to the technical field of biology and particularly relates to a method for removing keratan sulfate from a chondroitin sulfate crude extract. The method comprises the following steps of adding a sodium hydroxide solution into a chondroitin sulfate crude extract for reaction; then adding hydrochloric acid to regulate the PH value, filtering to obtain a filtrate, and adding sodium chloride into the filtrate to regulate the electrical conductivity of the solution to be 45-48ms / cm; then adding an ethanol solution, controlling the flow velocity of the ethanol solution to be 10-15mL / (L.min), precipitating chondroitin sulfate, and filtering to obtain a precipitate; dehydrating and drying the precipitate obtain a chondroitin sulfate finished product. The method is easy in control over technological conditions, low in cost, simple in operation and low in experimental apparatus requirement, and can be used for thoroughly solving the problems that removal of the keratan sulfate in chondroitin sulfate is difficult and industrial production is difficult to realize.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for removing keratan sulfate from chondroitin sulfate crude extract. Background technique [0002] Chondroitin Sulfate (CS) is a polymeric acidic mucopolysaccharide made from the cartilage of animals (pigs, cattle, horses, sheep, chickens, sharks, etc.), and its molecular structure is β-D-glucuronic acid and N- A polysaccharide chain composed of repeating disaccharide units formed from acetylgalactosamine. It is a white or slightly yellow powder with strong water absorption, easily soluble in water to form a viscous liquid, insoluble in organic solvents such as alcohol, acetone, ether and chloroform, and its salts are relatively stable to heat. [0003] Keratan sulfate (KS) is a kind of glycosaminoglycan, which is composed of different sulfated modified N-acetylglucosamine and D-galactose disaccharide repeating units. KS exists in the cornea of ​​animals in the form of prot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/08
CPCC08B37/0003C08B37/0069
Inventor 邢仕杰潘涛张富源殷晓薇
Owner RIZHAO LANSHAN BIOCHEM PROD
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