What are the principles and methods for detecting the sources of different species of heparin sodium

Currently, heparin sodium is mainly derived from the intestinal mucosa of pigs, cattle or sheep. However, heparin steel of bovine and sheep origin is at risk of being infected by related viruses, and the incidence of adverse reactions such as thrombocytopenia and thrombosis syndrome is much greater than that of pig-derived heparin steel. Therefore, in clinical use, people should try to choose porcine heparin sodium. However, in actual production, due to various reasons such as complex sources of production materials, the production raw materials from pigs are often contaminated by animal materials from cattle and sheep. Therefore, the establishment of methods for the identification of ingredients from pigs, sheep, cattle and other animal sources is essential to control the quality of drugs and prevent the contamination of xenogeneic animal-derived ingredients.

The following briefly talk about the detection principle of different species of heparin sodium, the detection principle: heparin sodium is a glycosaminoglycan, which is composed of uronic acid (L. iduronic acid, IdoA; D-glucose Uronic acid (GlcA) and glucosamine (6[.D-glucosamine, GlcN) are a mixture of polysaccharide chains with different chain lengths. Heparinase is used to enzymatically hydrolyze heparin sodium from different species, and its enzymatic hydrolysis products are analyzed and studied; that is, the species source of this product is characterized by the composition ratio of the enzymatic hydrolysis products (disaccharide mixture).

Heparinase is a type of polysaccharide lyase that acts on heparin or heparan sulfate molecules; there are three types of heparinase, namely heparinase I, heparinase II and heparinase III. Since each heparinase has a unique substrate specificity, they cut a specific site in the heparin chain. Heparinase 1 decomposes the Oc(1-4) glycosidic bond between GlcNs(6S) and IdoA(2S), and can cut off the antithrombin III binding pentasaccharide site in the heparin molecule; heparinase II can decompose two The sugar contains 2 or 3 sulfated groups of heparin, so its specificity is not strong; heparinase II1 decomposes the 6-position unsulfated GlcNS or IV. Heparin of Acetyl Glucosamine (GlcNAc) (see Figure 1). The proper mixing of the three heparinases can completely degrade heparin into an unsaturated disaccharide mixture with ultraviolet absorption at 234nm.

The detection of the source of heparin species mainly uses quantitative PCR to detect heparin sodium from different species, but this method is costly. The heparinase enzymatic hydrolysis method can effectively identify heparin samples mixed with sheep-derived crude heparin, and is simple and fast. The only disadvantage is that heparinase will lose the information of the uronic acid epimer during the enzymatic hydrolysis of heparin. Cannot distinguish between glucuronic acid and iduronic acid. In addition, in recent years, there have been methods for separating various disaccharides by capillary electrophoresis. Reversed-phase ion-pair chromatography can also be used to separate heparin disaccharides, and combined with mass spectrometry for quantitative analysis.

Desheng is a professional manufacturer of blood collection reagents and also has a complete detection system. The heparin sodium produced by it is mainly used as an additive for blood collection tubes. Heparin sodium has good anticoagulant effect, potency, and fast delivery.