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Method for separating and detecting over-sulfated chondroitin sulfate in heparin sodium

A technology of chondroitin polysulfate and a detection method, applied in the biological field, can solve problems such as ineffective detection, and achieve the effects of convenient quantitative determination, high resolution, and clear electrophoresis pattern

Active Publication Date: 2012-10-03
江苏麦德森制药有限公司
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Problems solved by technology

However, when the content of chondroitin polysulfate in heparin sodium is less than 0.5%, the method still cannot effectively detect

Method used

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  • Method for separating and detecting over-sulfated chondroitin sulfate in heparin sodium

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Embodiment Construction

[0025] The following is only an illustration of the present invention, rather than a limitation of the present invention.

[0026] (1) Separation and preparation of chondroitin polysulfate in heparin sodium:

[0027] a. Dissolving and filtering: Take 10 g of heparin sodium sample containing 0.2% polysulfate chondroitin by weight, add 150 mL of water and 5 g of sodium chloride into the mixed solution, heat up to 55 ° C ± 5 ° C, stir to completely dissolve the heparin sodium, and then use The weight concentration is 20% sodium hydroxide alkali solution to adjust the pH to 10-12, collect the filtrate by filtration, and cool the filtrate to zero, adjust the pH to 1-2 with hydrochloric acid, and collect the filtrate by filtration;

[0028] b, rapid precipitation: the filtrate is adjusted to PH to 5.5 with sodium hydroxide, and 95% alcohol is slowly added under stirring, so that the weight concentration of the solution containing alcohol is 35%, and it is allowed to stand still; use...

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Abstract

The invention discloses a method for separating and detecting over-sulfated chondroitin sulfate in heparin sodium. The method uses a fast precipitation method to separate the over-sulfated chondroitin sulfate from the heparin sodium, and further detect an object obtained after the separation by using agarose and electrophoresis to realize the detection of the low-content over-sulfated chondroitinsulfate in the heparin sodium, wherein the sensibility reaches over 0.1 percent. The electrophoresis process and result can be directly detected and observed by using an ultraviolet lamp, and the observation is intuitive and is easy to judge. A zone after the electrophoresis is easy to be dyed, a sample can be eluted very easily to provide convenience for quantitative determination, and the sample can be preserved for a long time after being prepared into a dry film.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a pretreatment and electrophoresis detection method of chondroitin polysulfate in heparin sodium. Background technique [0002] my country is the country with the largest output of heparin sodium, and more than 70% of the raw materials of heparin sodium products in the international market come from China. With the increasing demand, the price of heparin sodium has been rising all the way in recent years. In February 2008, 4 deaths occurred in the United States due to the injection of heparin sodium, and there were many cases of medical accidents with adverse reactions, which became the "heparin sodium incident" that shocked the world. According to the analysis of relevant experts, the above-mentioned accident may be related to the polysulfate chondroitin contained in heparin sodium. The Chinese government attaches great importance to and pays close attention to the development of th...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N1/28G01N1/34G01N1/30
Inventor 刘榜惠张志斌袁红英
Owner 江苏麦德森制药有限公司
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