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Method for detecting Sulodexide with enzymolysis-HPLC

A sulodexide enzymatic and enzymatic hydrolysis technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of short polysaccharide chain length and low sulfation degree

Pending Publication Date: 2017-07-07
SHENZHEN HEPALINK PHARMA GRP CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The LMWH part contains the same disaccharide polymer structure as unfractionated heparin, but the degree of sulfation is lower and the length of the polysaccharide chain is shorter

Method used

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  • Method for detecting Sulodexide with enzymolysis-HPLC
  • Method for detecting Sulodexide with enzymolysis-HPLC
  • Method for detecting Sulodexide with enzymolysis-HPLC

Examples

Experimental program
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Embodiment 1

[0022] a, the preparation of the solution in the complete enzymatic hydrolysis step of sulodexide:

[0023] Preparation of buffer solution A: Weigh 68 mg of potassium dihydrogen phosphate, dissolve it with about 30 ml of ultrapure water, neutralize it with 1 mol / L sodium hydroxide solution to pH 7, add 10 mg of bovine albumin, and transfer to In a 50ml volumetric flask, make up to volume with ultrapure water and shake well.

[0024] Preparation of buffer B: pipette 0.58ml of glacial acetic acid into a beaker, add about 80ml of ultrapure water, add 32mg of calcium acetate, and dissolve it. Neutralize to pH 7 with 1mol / L sodium hydroxide solution, and add 10 mg of bovine albumin. Move to a 100ml volumetric flask, make up to volume with ultrapure water, and shake well. A 100 mM solution of sodium acetate was obtained.

[0025] Preparation of heparinase solution: Dilute the heparinase to 0.4IU / ml with buffer A according to the amount of heparinase I, heparinase II, and heparina...

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Abstract

The invention discloses a method for detecting Sulodexide with enzymolysis-HPLC. The method includes the following steps of a, complete enzymolysis of Sulodexide, wherein a mixed enzyme solution of an enzyme I, an enzyme II and an enzyme III is added into a Sulodexide solution for enzymolysis at the room temperature; b, HPLC analysis, wherein a degradation product is taken for SAX-HPLC analysis; c, calculation of the types and contents of disaccharide units, wherein the types and contents of test sample heparin disaccharide units are determined according to the elution time of eight existing standard heparin disaccharides, and the percentage content of each disaccharide is calculated.

Description

technical field [0001] The invention relates to a method for detecting sulodexide raw materials, in particular to a method for confirming the chemical composition of sulodexide by analyzing the degradation products by SAX-HPLC after combined degradation with heparanase. Background technique [0002] Sulodexide, whose trade name is VESSEL DUE F, and whose chemical name is glucuronyl glucosaminoglycan sulfate. Originally produced by Alfa Wassermann in Italy, it was first launched in Italy in 1974 and entered the Chinese market in 2003. So far, only the original research company has sales in China. The indications are vascular diseases with the risk of thrombosis, and it is an orally absorbable anticoagulant drug. In recent years, it has been shown that it is effective in the treatment of diabetic nephropathy. [0003] Sulodexide is extracted from pig intestinal mucosa. It is a highly purified glycosaminoglycan. It is a long-chain negatively charged macromolecule composed of...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 马小来李浩杰张紫恒李锂
Owner SHENZHEN HEPALINK PHARMA GRP CO LTD
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