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

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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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 identify the structure of the intact sugar chain of low molecular weight heparin

[0004] In addition to the methods mentioned above, the detection methods for low molecular weight heparin stipulated in the United States Pharmacopoeia, European Pharmacopoeia and Chinese Pharmacopoeia are also for the analysis of low molecular weight heparin mixtures, which can only characterize the mixture, but not the individual components. direct analysis

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