The principle and method of detecting the source of heparin sodium?

  At present, heparin mainly comes from the intestinal mucosa of pigs, cattle and sheep. However, bovine and sheep heparin steels are at risk of infection with related viruses, and the incidence of adverse events such as low platelet count and thrombotic syndrome is much higher than that of pig heparin steels. Therefore, porcine heparin sodium should be used as far as possible in clinical application. However, in actual production, due to the complex sources of raw materials and various reasons, the production of raw materials from pigs, cattle, sheep and other sources of animal raw materials are often contaminated. Therefore, it is of great significance to establish a method for identification of pig, sheep and cattle sources to control the quality of drugs and prevent contamination from different animal sources.

  Let me just talk a little bit about how different kinds of heparin are tested. Detection principle: Heparin sodium is a glycosaminoglycan composed of uronic acid (L. iduronic acid, IDOad-glucuronic acid, GlcA) and glucosamine (6[. D-glucosamine, GlcN). Different kinds of heparin sodium are hydrolyzed with heparin enzyme, and their hydrolyzed products are analyzed and studied, that is, The type of source of this product is characterized by the composition of the hydrolysate (the disaccharide mixture).

The principle and method of detecting the source of heparin sodium?

  Heparinase is a polysaccharide lyase that acts on heparin or heparan sulfate. There are three types of heparinase: heparinase I, heparinase II and heparinase III. Because each heparin enzyme has a unique stroma specificity, they can cut specific sites on the heparin chain. Heparin 1 destroys the glucoside bond between GLCNS (6s) and IDOA (2s), cutting off the pentagose site in the heparin molecule where antithrombin III binds, while heparin II destroys two or three sulfating groups in heparin and disaccharides. Heparinase II1 destroys gICNS or IV containing six acidification sites. Glucosamine in heparin (see Figure 1). When properly mixed with the three kinds of heparin enzymes, heparin can be completely degraded into the unsaturated disaccharide mixture, whose UV absorption wavelength is 234nm.

  The detection of heparin species is mainly to identify heparin sodium from different species by quantitative polymerase chain reaction, but this method is costly. Heparin enzyme hydrolysis can effectively identify heparin samples mixed with crude heparin from sheep, and it is simple and fast. The only disadvantage of heparinase is that the information of the differential isomer of the uronic acid is lost during the enzymatic hydrolysis of heparin and the glucuronic acid and iduronic acid cannot be distinguished. In addition, in recent years, capillary electrophoresis has been used to separate various disaccharides. Heparin disaccharides can also be separated by reversed-phase ion pair chromatography and quantified by mass spectrometry.

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