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Comprehensive Identification and Quantitation of Basic Building Blocks for Low-Molecular Weight Heparin
Xiaojun Sun1,2, Anran Sheng1, Xinyue Liu1,2
1National Glycoengineering Research Center, Shandong Provincial Key Laboratory of Carbohydrate Chemistry and Glycobiology, and State Key Laboratory of Microbial Technology, Shandong University , Jinan, Shandong 250100, China.
Analytical Chemistry
|July 9, 2016
Summary
This study presents a new analytical method to characterize low-molecular weight heparins (LMWHs). The method precisely identifies LMWH building blocks, ensuring drug quality and safety.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Pharmacology
Background:
- Low-molecular weight heparins (LMWHs) are crucial anticoagulants derived from heparin.
- Chemical depolymerization and side reactions create complex LMWH structures.
- Precise structural characterization is vital for LMWH drug quality and safety.
Purpose of the Study:
- To develop a comprehensive analytical method for characterizing LMWH structures.
- To identify and quantify the basic building blocks of LMWHs.
- To ensure the quality and safety of LMWH anticoagulant drugs.
Main Methods:
- Combined enzymatic digestion and oxidative degradation of LMWHs.
- Hydrophilic interaction chromatography coupled with triple quadrupole mass spectrometry (HILIC-MS/MS).
- Multiple reaction monitoring (MRM) mode for sensitive detection and quantitation.
Main Results:
- Over 30 natural and chemically modified heparin building blocks were detected after enzymatic digestion.
- Two novel structures, a trisaccharide and a tetrasaccharide, were identified.
- Analysis of reduced and oxidatively degraded samples provided information on LMWH termini.
- The method demonstrated reproducibility and was applied to enoxaparin samples.
Conclusions:
- The developed method enables detailed characterization of LMWH structures.
- It facilitates the identification of novel heparin structures.
- The method is applicable for assessing the sameness of LMWH products, ensuring drug consistency.
