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Method for qualitative and quantitative analysis of pentasaccharide structure units in enoxaparin sodium

A technology of enoxaparin sodium and structural units, which is applied in the direction of material analysis, analysis materials, and measuring devices by electromagnetic means, can solve the problem that the qualitative and quantitative determination of pentasaccharide structural units cannot be realized, and the quality control of enoxaparin sodium cannot be satisfied. Requirements and other issues, to achieve the effect of significant industrial application value, scientific and convenient analysis means

Inactive Publication Date: 2017-10-20
SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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Problems solved by technology

Although this method can be used to identify the disaccharides, trisaccharides, tetrasaccharides and oligosaccharides with 1,6-cycloside structure in enoxaparin sodium, but this method cannot realize the qualitative and quantitative analysis of the pentasaccharide structural units therein. Quantitative determination, so the complete qualitative and quantitative analysis of the fine structure in enoxaparin sodium cannot be truly realized, and the quality control requirements of enoxaparin sodium cannot be met

Method used

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  • Method for qualitative and quantitative analysis of pentasaccharide structure units in enoxaparin sodium
  • Method for qualitative and quantitative analysis of pentasaccharide structure units in enoxaparin sodium
  • Method for qualitative and quantitative analysis of pentasaccharide structure units in enoxaparin sodium

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Embodiment

[0054] 1. Preparation of sample solution

[0055] (1) Preparation of incubation buffer solution: Weigh 840.07mg of sodium bicarbonate and 1.461g of sodium chloride in 40mL of water, adjust the pH to 8.3 with 1M hydrochloric acid solution, transfer the solution to a 50mL volumetric flask, and dilute to Scale; Before use, filter with a filter membrane with a pore size of 0.45 μm.

[0056] (2) Preparation of binding buffer solution: Weigh 302.85mg tris(hydroxymethyl)aminomethane and 2.1915g sodium chloride in 230mL water, adjust to pH 7.4 with 1M hydrochloric acid solution, then transfer the solution to a 250mL volumetric flask In the medium, make the volume to the mark; before use, filter with a filter membrane with a pore size of 0.45 μm.

[0057] (3) Preparation of elution buffer solution: Weigh 302.85mg of tris(hydroxymethyl)aminomethane and 29.22g of sodium chloride in 230mL of water, adjust the pH to 7.4 with 1M hydrochloric acid solution, and transfer the solution to a ca...

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Abstract

The invention discloses a method for qualitative and quantitative analysis of pentasaccharide structure units in enoxaparin sodium. The method comprises the steps as follows: a) an enoxaparin sodium sample is separated with an affinity chromatographic column of antithrombase III, and multiple different affinity active components are prepared; b) the enoxaparin sodium sample and the multiple different affinity active components are subjected to complete enzymolysis with mixed heparinase; c) each completely enzymatic hydrolysate is subjected to capillary electrophoresis separation analysis; d) the electrophoresis mobility of each separated component is calculated according to the formula shown in the specification; e) the enzymatic hydrolysate corresponding to the separation peak in a capillary electrophoretogram of each completely enzymatic hydrolysate is authenticated in combination with the linear relation, shown in the specification, between the charge mass ratio and the electrophoresis mobility of oligosaccharide, so that separation peaks of enzymatic fragments of the pentasaccharide structure units are qualitatively determined; f) the content of the pentasaccharide structure units is quantitatively calculated according to peak areas of the separation peaks of the determined enzymatic fragments of the pentasaccharide structure units.

Description

technical field [0001] The invention relates to a method capable of qualitatively and quantitatively analyzing the pentasaccharide structural unit in enoxaparin sodium, and belongs to the technical fields of analytical chemistry and drug analysis. Background technique [0002] Heparin is a disaccharide repeating unit composed of uronic acid (L-iduronic acid IdoA; D-glucuronic acid, GlcA) and glucosamine (α-D glucosamine, GlcN) with different chain lengths. A class of linear sulfated mucopolysaccharides with good anticoagulant and antithrombotic properties, used in the treatment of venous thromboembolism induced after surgery. Enoxaparin sodium is a low-molecular-weight heparin produced by heparin extracted from pig intestinal mucosa, which is first esterified to obtain heparin sodium benzyl ester derivatives, and then cleaved by β-elimination under alkaline conditions. The structure of enoxaparin sodium is more complex due to the differences in the sulfation site and degree...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 康经武张明瑜
Owner SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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