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Method for detecting complete degradation product of low molecular weight heparin based on post-column derivatization

A low-molecular-weight heparin, detection method technology, applied in the direction of measurement device, material separation, analysis material, etc., can solve the problems affecting the accuracy and repeatability of analysis, chromatographic separation interference, affecting analysis results, etc., to achieve great practical value, analysis High degree of automation and good repeatability

Active Publication Date: 2014-06-11
SUZHOU KANGHESHUN MEDICAL TECH
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Problems solved by technology

The disadvantage of this method is that the operation process is cumbersome, and it is easy to affect the accuracy and repeatability of the analysis. The by-products of the derivatization reaction may cause great interference to the chromatographic separation and affect the analysis results.

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  • Method for detecting complete degradation product of low molecular weight heparin based on post-column derivatization
  • Method for detecting complete degradation product of low molecular weight heparin based on post-column derivatization
  • Method for detecting complete degradation product of low molecular weight heparin based on post-column derivatization

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Embodiment Construction

[0025] The following will clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0026] A method for detecting a complete enzymatic hydrolysis product of low molecular weight heparin is provided, comprising the steps of:

[0027] (1) Low molecular weight heparin sodium was degraded with heparinase I, II and III at 25°C for 48 h, the degradation product was filtered by ultrafiltration membrane, dried under vacuum and reduced pressure, and then prepared into 1 mg / mL preparation with deionized water. test solution.

[0028] (2) Preparation of the first mobile phase: Dissolve tetrabutylammonium bisu...

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Abstract

The invention discloses a method for detecting a complete degradation product of low molecular weight heparin based on post-column derivatization. The method comprises the following steps: degrading low molecular weight heparin to obtain a complete enzymatic product, separating the product by virtue of ion pair reversion phase chromatography, further carrying out post-column fluorescence derivation reaction, subsequently carrying out fluorescence detection, and analyzing to obtain the complete degradation product. In such a mode, the method of ion pair reversion phase chromatography with the combination of post-column fluorescence derivation is adopted, both detectable fluorescent groups are provided for low molecular weight heparin without ultraviolet absorption, and fluorescence detection with high sensitivity is achieved, and great practical values are achieved for development, production control and medicine security guarantee in low molecular weight heparin medicines.

Description

technical field [0001] The invention relates to a detection method combining ion-pair reversed-phase chromatography and post-column derivatization of complete enzymatic hydrolysis products of low-molecular-weight heparin, and belongs to the technical field of medicine, raw material medicine and raw material detection. Background technique [0002] Heparin is a highly sulfated linear glycosaminoglycan that widely exists in animal organs and tissues and plays an important role in biological activities and physiological processes by binding to many proteins. Heparin has biological functions such as anticoagulation, anti-inflammation, anti-allergy, anti-virus, anti-cancer, etc., and as a natural anti-coagulant and anti-thrombotic drug, it is widely used in clinical disease treatment, but in the long-term use process, it will cause Cause side effects such as osteoporosis and thrombocytopenia. Low-molecular-weight heparin is a fragment with a smaller molecular weight obtained by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 白桦
Owner SUZHOU KANGHESHUN MEDICAL TECH
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