Ion pair antiphase chromatography and mass spectrometry combined detection method for low-molecule heparin part degradation products

A low-molecular-weight heparin and detection method technology, which is applied in the detection field of ion-pair reversed-phase chromatography and high-resolution mass spectrometry, can solve the problems of enhancing the heterogeneity of enzymatic hydrolysis products, incapable of accurate qualitative analysis of sugar chain fragments, and difficult analysis.

Active Publication Date: 2013-12-18
HEBEI CHANGSHAN BIOCHEM PHARMA
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AI Technical Summary

Problems solved by technology

Fingerprint analysis of partially degraded fragments of low-molecular-weight heparin is a very important link in its structural characterization, through which the sequence information of heparin sugar chains can be obtained, but partial degradation of low-molecular-weight heparin by heparinase will enhance the heterogeneity of enzymatic hydrolysis products , making it more difficult to analyze
Generally, method

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  • Ion pair antiphase chromatography and mass spectrometry combined detection method for low-molecule heparin part degradation products
  • Ion pair antiphase chromatography and mass spectrometry combined detection method for low-molecule heparin part degradation products
  • Ion pair antiphase chromatography and mass spectrometry combined detection method for low-molecule heparin part degradation products

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Embodiment Construction

[0005] A method for detecting partial degradation products of low-molecular-weight heparin by ion-pair reversed-phase chromatography-mass spectrometry, the steps of which are as follows:

[0006] (1) Dissolve the amine ion-pair reagent in deionized water to prepare mobile phase A with an amine ion-pair concentration of 10-40mM, and adjust the pH to 5.5-8.5;

[0007] Alternatively, dissolving the amine ion-pair reagent and hexafluoroisopropanol in deionized water to prepare a mobile phase A' with an amine ion-pair concentration of 10-40 mM and a hexafluoroisopropanol concentration of 20-100 mM;

[0008] (2) Dissolve the amine ion pair reagent in 75% by volume acetonitrile or 75% by volume methanol solution to prepare mobile phase B with an amine ion pair concentration of 10-40mM, and adjust the pH to 5.5~8.5;

[0009] Alternatively, the amine ion pair reagent and hexafluoroisopropanol are dissolved in 75% by volume acetonitrile or 75% by volume methanol solution to obtain an a...

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Abstract

The invention relates to an ion pair antiphase chromatography and mass spectrometry combined detection method for low-molecule heparin part degradation products. All components of the low-molecule heparin part degradation products are detected in a manner of combining ion pair antiphase chromatography and high-resolution mass spectrometer; all the components in a sample are separated through the ion pair antiphase chromatography, and a precise molecule weight is obtained through the high-resolution mass spectrometer, so that sugar chain sequence information, including structures of two ends, lengths of sugar chains and substitution numbers of acetyl and sulfuric acid, of all the components is calculated, so that the structures of the low-molecule heparin part degradation products can be finely represented. The ion pair antiphase chromatography and mass spectrometry combined detection method can detect the structures of the two ends, the lengths of the sugar chains and the substitution numbers of the acetyl and the sulfuric acid of the low-molecule heparin part degradation products; the problem in the prior art that only an ultraviolet absorption atlas for degradation of low-molecule heparin parts can be obtained is solved; the ion pair antiphase chromatography and mass spectrometry combined detection method has a high practicability value in improvement of the detection level of heparin and guarantee of drug safety.

Description

technical field [0001] The invention relates to an ion-pair reversed-phase chromatography and high-resolution mass spectrometry combined detection method for partially degraded 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 glycosaminoglycan anticoagulant and antithrombotic drug. Low molecular weight heparin is a new type of antithrombotic drug prepared by degrading heparin by physical, chemical or enzymatic methods. It has strong polarity, high heterogeneity, It is difficult to carry out structural characterization due to the instability of the sulfuric acid group. Fingerprint analysis of partially degraded fragments of low-molecular-weight heparin is a very important link in its structural characterization, through which the sequence information of heparin sugar chains can be obtained, but partial degradation of low-molecular-weight hepari...

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Application Information

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IPC IPC(8): G01N30/89
Inventor 迟连利徐晓晖李道远孙晓君
Owner HEBEI CHANGSHAN BIOCHEM PHARMA
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