PMID- 25701653 OWN - NLM STAT- MEDLINE DCOM- 20151229 LR - 20150406 IS - 1873-426X (Electronic) IS - 0008-6215 (Linking) VI - 407 DP - 2015 Apr 30 TI - Fragment profiling of low molecular weight heparins using reversed phase ion pair liquid chromatography-electrospray mass spectrometry. PG - 26-33 LID - S0008-6215(15)00029-4 [pii] LID - 10.1016/j.carres.2015.01.016 [doi] AB - Low molecular weight heparins (LMWHs) are linear and highly charged carbohydrate polymers prepared by chemical or enzymatic depolymerization of heparin. Compared to unfractionated heparin (UFH), LMWHs are prevalently used as clinical anticoagulant drugs due to their lower side effects and better bioavailability. The work presented herein provides a rapid and powerful fragment mapping method for structural characterization of LMWHs. The chain fragments of two types of LMWHs, enoxaparin and nadroparin, were generated by controlled enzymatic digestion with each of heparinase I (Hep I, Enzyme Commission (EC) # 4.2.2.7), heparinase II (Hep II, no EC # assigned) and heparinase III (Hep III, EC # 4.2.2.8). Reversed phase ion pair high performance liquid chromatography (RPIP-HPLC) coupled with electrospray ion trap time-of-flight mass spectrometry (ESI-IT-TOF-MS) was used to profile the oligosaccharide chains ranging from disaccharides to decasaccharides. A database containing all theoretical structural compositions was established to assist the mass spectra interpretation. The six digests derived by three enzymes from two types of LMWHs exhibited distinguishable fingerprinting patterns. And a total of 94 enoxaparin fragments and 109 nadroparin fragments were detected and identified. Besides the common LMWH oligosaccharides, many components containing characteristic LMWH structures such as saturated L-idopyranosuronic acid, 2,5-anhydro-D-mannitol, 1,6-anhydro-D-aminopyranose, as well as odd number oligosaccharides were also revealed. Quantitative comparison of major components derived from innovator and generic nadroparin products was presented. This approach to profile LMWHs' fragments offers a highly reproducible, high resolution and information-rich tool for evaluating the quality of this category of anticoagulant drugs or comparing structural similarities among samples from various sources. CI - Copyright (c) 2015 Elsevier Ltd. All rights reserved. FAU - Xu, Xiaohui AU - Xu X AD - National Glycoengineering Research Center, Shandong University, Jinan 250100, China; State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China. FAU - Li, Daoyuan AU - Li D AD - National Glycoengineering Research Center, Shandong University, Jinan 250100, China; State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China. FAU - Chi, Lequan AU - Chi L AD - School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY 11794, USA. FAU - Du, Xuzhao AU - Du X AD - Hebei Changshan Biochemical Pharmaceutical Co. Ltd., Shijiazhuang 050800, China. FAU - Bai, Xue AU - Bai X AD - National Glycoengineering Research Center, Shandong University, Jinan 250100, China. FAU - Chi, Lianli AU - Chi L AD - National Glycoengineering Research Center, Shandong University, Jinan 250100, China; State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China. Electronic address: lianlichi@sdu.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150202 PL - Netherlands TA - Carbohydr Res JT - Carbohydrate research JID - 0043535 RN - 0 (Anticoagulants) RN - 0 (Enoxaparin) RN - 0 (Heparin, Low-Molecular-Weight) RN - 0 (Nadroparin) RN - EC 4.2.2.- (Polysaccharide-Lyases) RN - EC 4.2.2.- (heparinase II) RN - EC 4.2.2.7 (Heparin Lyase) RN - EC 4.2.2.8 (heparitinsulfate lyase) SB - IM MH - Anticoagulants/*chemistry/isolation & purification MH - Carbohydrate Sequence MH - Chromatography, Reverse-Phase/*methods MH - Databases, Chemical MH - Enoxaparin/chemistry/isolation & purification MH - Heparin Lyase/metabolism MH - Heparin, Low-Molecular-Weight/*chemistry/isolation & purification MH - Nadroparin/chemistry/isolation & purification MH - Polysaccharide-Lyases/metabolism MH - Spectrometry, Mass, Electrospray Ionization/*methods OTO - NOTNLM OT - Fragment profiling OT - Low molecular weight heparins OT - Mass spectrometry OT - Reversed phase ion pair EDAT- 2015/02/24 06:00 MHDA- 2015/12/30 06:00 CRDT- 2015/02/22 06:00 PHST- 2014/09/29 00:00 [received] PHST- 2014/12/12 00:00 [revised] PHST- 2015/01/23 00:00 [accepted] PHST- 2015/02/22 06:00 [entrez] PHST- 2015/02/24 06:00 [pubmed] PHST- 2015/12/30 06:00 [medline] AID - S0008-6215(15)00029-4 [pii] AID - 10.1016/j.carres.2015.01.016 [doi] PST - ppublish SO - Carbohydr Res. 2015 Apr 30;407:26-33. doi: 10.1016/j.carres.2015.01.016. Epub 2015 Feb 2.