期刊
ANALYTICAL BIOCHEMISTRY
卷 451, 期 -, 页码 35-41出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2014.02.005
关键词
LMWH; DAD; AMAC derivatization; ESI-MS; Compositional analysis
资金
- National Major Scientific and Technological Special Project for Significant New Drugs Development [2012ZX0950200-005]
- National Basic Research Program of China (973 Program) [2012CB822102]
- National Natural Science Foundation of China [31000367]
- Natural Science Foundation of Shandong Province, China [ZR2011CM038, ZR2010CM041]
Low molecular weight heparins (LMWHs) are important artificial preparations from heparin polysaccharide and are widely used as anticoagulant drugs. To analyze the structure and composition of LMWHs, identification and quantitation of their natural and modified building blocks are indispensable. We have established a novel reversed-phase high-performance liquid chromatography-diode array detection-electrospray ionization-mass spectrometry approach for compositional analysis of LMWHs. After being exhaustively digested and labeled with 2-aminoacridone, the structural motifs constructing LMWHs, including 17 components from dalteparin and 15 components from enoxaparin, were well separated, identified, and quantified. Besides the eight natural heparin disaccharides, many characteristic structures from dalteparin and enoxaparin, such as modified structures from the reducing end and nonreducing end, 3-O-sulfated tetrasaccharides, and trisaccharides, have been unambiguously identified based on their retention time and mass spectra. Compared with the traditional heparin compositional analysis methods, the approach described here is not only robust but also comprehensive because it is capable of identifying and quantifying nearly all components from lyase digests of LMWHs. (C) 2014 Elsevier Inc. All rights reserved.
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