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Method for precisely and quantitatively controlling chondroitin sulfate and dermatan sulfate contents of heparin/heparinoid

A technology of dermatan sulfate and chondroitin sulfate, applied in the field of precise and quantitative control of chondroitin sulfate and dermatan sulfate content in heparin/heparin, can solve the problems affecting the safety of heparin product use and small production scale

Inactive Publication Date: 2015-06-24
TSINGHUA UNIV
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Problems solved by technology

However, the production scale of this type of heparin product is small, the main reason is that there is no effective means to precisely control the content of chondroitin sulfate and dermatan sulfate, and the difference in the amount of these impurities will significantly affect the quality of this type of heparin product. Efficacy and even safety

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  • Method for precisely and quantitatively controlling chondroitin sulfate and dermatan sulfate contents of heparin/heparinoid
  • Method for precisely and quantitatively controlling chondroitin sulfate and dermatan sulfate contents of heparin/heparinoid
  • Method for precisely and quantitatively controlling chondroitin sulfate and dermatan sulfate contents of heparin/heparinoid

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Embodiment 1

[0091] Embodiment 1, refined danaparin sodium sample

[0092] 1. Content analysis method

[0093] 1.1 DS content analysis method: prepare the test sample solution 10mg / ml with Tris buffer solution of pH 7.4, take 1ml of the sample solution, add 1IU of MBP-ChSase B at 25°C for 24h, and then use HPLC analysis (specific chromatogram The conditions are as follows), and the DS content was calculated by the external standard curve. The detection limit of DS content is 0.01%. Compared with the DS content analysis method recorded in European Pharmacopoeia 7.0 (EP7.0), the HPLC analysis method has high sensitivity and good repeatability.

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Abstract

The invention discloses a method for precisely and quantitatively controlling the chondroitin sulfate and dermatan sulfate contents of heparin / heparinoid. The method comprises the following steps: 1) adding proteins with chondrosulphatase B activity into a heparinoid / heparin containing solution, and after the DS content reaches an expected value, terminating the enzymatic hydrolysis; and 2) further adding proteins with chondrosulphatase AC activity into the solution, and after the CS content reaches an expected value, terminating the enzymatic hydrolysis, thus obtaining a heparin sample or heparinoid sample with expected CS and DS contents. The method disclosed by the invention realizes the efficient and precise control over the CS and DS contents of heparin / heparinoid, thereby establishing a novel low-cost and high-yield production process of heparin products. The method is simple and easy to perform, has an application value of large-scale production, and can be used for producing danaparoid sodium meeting the standards of European Pharmacopoeia.

Description

technical field [0001] The invention relates to a method for precisely quantitatively controlling the content of chondroitin sulfate and dermatan sulfate in heparin / heparin. Background technique [0002] 1.1 Heparin [0003] 1.1.1 Introduction to Heparin: [0004] Heparin (Heparin, HP) was first named after the discovery of the liver. Heparin is widely distributed in mammalian tissues, such as liver, lung, mesentery, heart, spleen, kidney, thymus, placenta, muscle and blood (high Lighting, Cao Xianping, Zhang Yubing, Analysis of mucopolysaccharide composition in porcine small intestinal mucosa / serosa / casing [J], Chemical and Bioengineering, 2012, 29(4): 91-94). The content of heparin in various tissues is related to the number of mast cells. The granules in mast cells contain heparin or heparin precursors. When physical or chemical stimuli degranulate mast cells, heparin is released and is produced by the liver in vivo. It is excreted from the urine due to the inactivatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/10
Inventor 邢新会苏楠张翀梅祥
Owner TSINGHUA UNIV
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