Method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum

A low-molecular-weight heparin and detection method technology, which is applied in material separation, measurement devices, and analytical materials, etc., can solve problems such as the inability to analyze a single component and the inability to identify the sugar chain structure of low-molecular-weight heparin

Active Publication Date: 2012-10-31
HEBEI CHANGSHAN BIOCHEM PHARMA
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Problems solved by technology

Using heparinase to degrade low-molecular-weight heparin into heparin disaccharides, and then using ion-pair reversed-phase chromatography and mass spectrometry to analyze the composition of disaccharides is a commonly used characterization method in recent years (Zhang F, et al. Analytical and Bioanalytical Chemistry, 2011,401(9):2793-2803.), but because the detection sample is the degraded heparin disaccharide, this method cannot id

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  • Method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum
  • Method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum
  • Method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum

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[0007] A low-molecular-weight heparin ion-pair reversed-phase chromatography-mass spectrometry detection method, the steps are as follows:

[0008] (1) Dissolve the amine ion pair reagent in deionized water, adjust the pH value to 5.5-8.5 with a pH adjusting reagent, and prepare mobile phase A with a concentration of 10-30mM; the concentration mentioned in this step refers to the concentration of amine ions The molar concentration of the solute in mobile phase A in the reagent;

[0009] (2) Dissolve the amine ion-pair reagent in 75% by volume acetonitrile or 75% by volume methanol solution, adjust the pH value to 5.5-8.5 with a pH adjustment reagent, and prepare mobile phase B with a concentration of 10-30mM Concentration described in this step refers to the molar concentration of solute in mobile phase B in the amine ion pair reagent;

[0010] (3) Dissolve the low-molecular-weight heparin sample to be tested in mobile phase A to prepare a test solution with a concentration o...

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Abstract

The invention relates to a method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum. The method comprises the steps of: detecting all compositions of low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and high-resolution mass spectrum, separating main compositions from a sample through the ion pair reversed phase chronmatogaphy, obtaining precise molecular weight by the high-resolution mass spectrum, and calculating information of a carbohydrate chain sequence, including structures of two ends, length of a carbohydrate chain and substitution number of ethanoyl and sulfate, thus carrying out fine surface features on the low-molecular-weight heparin. The method has great practical values on improving the heparin detection level and ensuring drug safety.

Description

technical field [0001] The invention relates to a low-molecular-weight heparin detection method combined with ion-pair reversed-phase chromatography and high-resolution mass spectrometry, and belongs to the technical field of medicine, raw material medicine and raw material detection. Background technique [0002] Heparin is a glycosaminoglycan drug with strong anticoagulant effect and is the drug of choice for the treatment of thromboembolic diseases. Low-molecular-weight heparin is prepared by chemically or enzymatically degrading heparin, and common ones include enoxaparin sodium (Enoxaparin), nadroparin calcium (Nadroparin) and dalteparin sodium (Dalteparin). Compared with heparin, low-molecular-weight heparin has higher antithrombotic activity, lower anticoagulant activity, less bleeding side effects, high bioavailability, and long half-life in vivo. It has received more and more attention and application. [0003] Because heparin and low molecular weight heparin have ...

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Application Information

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IPC IPC(8): G01N30/89
Inventor 迟连利李道远王章杰
Owner HEBEI CHANGSHAN BIOCHEM PHARMA
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