Bottom-up low molecular weight heparin analysis using liquid chromatography-Fourier transform mass spectrometry for extensive characterization.
Bottom-up low molecular weight heparin analysis using liquid chromatography-Fourier transform mass spectrometry for extensive characterization.
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DOI：
10.1021/ac501301v
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发表时间：
2014-07-01
影响因子：
7.4
通讯作者：
Li, Lingyun
中科院分区：
文献类型：
--
作者：
Li, Guoyun;Steppich, Julia;Wang, Zhenyu;Sun, Yi;Xue, Changhu;Linhardt, Robert J.;Li, Lingyun
低分子量肝素（LMWH）是一种多相、多分散、带高度负电荷的糖胺聚糖链混合物，可用作抗凝剂。LMWH的详细表征对于药物质量保证和新药研发具有重要意义。在这项研究中，在线亲水相互作用色谱（HILIC）傅立叶变换质谱（FTMS）分析低分子量肝素裂解酶II消化产生的寡糖片段。对40多种低分子量肝素寡糖片段进行了定量，并用于比较三个不同制造商制备的低分子量肝素。定量的片段结构包括来自这些LMWH的主要重复单元的不饱和二糖/寡糖、来自其抗凝血酶III结合位点的含3-O-磺基的四糖、在其生产过程中形成的含1，6-脱水环的寡糖、来自一些链非还原末端的饱和糖醛酸寡糖和来自一些链还原末端的氧化连接区寡糖。这种自下而上的方法提供了丰富详细的结构分析和定量信息，具有高准确性和可重复性。当与自上而下的方法相结合时，基于HILIC LC-FTMS的分析应适用于LMWH生产中的先进质量控制和质量保证。
Low molecular weight heparins (LMWHs) are heterogeneous, polydisperse, and highly negatively charged mixtures of glycosaminoglycan chains prescribed as anticoagulants. The detailed characterization of LMWH is important for the drug quality assurance and for new drug research and development. In this study, online hydrophilic interaction chromatography (HILIC) Fourier transform mass spectrometry (FTMS) was applied to analyze the oligosaccharide fragments of LMWHs generated by heparin lyase II digestion. More than 40 oligosaccharide fragments of LMWH were quantified and used to compare LMWHs prepared by three different manufacturers. The quantified fragment structures included unsaturated disaccharides/oligosaccharides arising from the prominent repeating units of these LMWHs, 3-O-sulfo containing tetrasaccharides arising from their antithrombin III binding sites, 1,6-anhydro ring-containing oligosaccharides formed during their manufacture, saturated uronic acid oligosaccharides coming from some chain nonreducing ends, and oxidized linkage region oligosaccharides coming from some chain reducing ends. This bottom-up approach provides rich detailed structural analysis and quantitative information with high accuracy and reproducibility. When combined with the top-down approach, HILIC LC-FTMS based analysis should be suitable for the advanced quality control and quality assurance in LMWH production.
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影响因子：
7.3
作者：
Guerrini, Marco;Elli, Stefano;Viskov, Christian
通讯作者：
Viskov, Christian
影响因子：
7.4
作者：
Brustkern, Adam M.;Buhse, Lucinda F.;Keire, David A.
通讯作者：
Keire, David A.
影响因子：
4.1
作者：
Galeotti, Fabio;Volpi, Nicola
通讯作者：
Volpi, Nicola
影响因子：
7.4
作者：
Li, Lingyun;Zhang, Fuming;Zaia, Joseph;Linhardt, Robert J.
通讯作者：
Linhardt, Robert J.
影响因子：
3.1
作者：
Keire, David A.;Buhse, Lucinda F.;Al-Hakim, Ali
通讯作者：
Al-Hakim, Ali