Ion-pairing liquid chromatography with on-line electrospray ion trap mass spectrometry for the structural analysis of N-unsubstituted heparin/heparan sulfate.
The rare N-unsubstituted glucosamine (GlcNH3(+)) residues in heparan sulfate (HS) have important biological and pathophysiological roles. In this study, a high-resolution method for the separation and analysis of N-unsubstituted disaccharides of heparin/HS is described. Four N-unsubstituted disaccharides, together with eight N-substituted species, can be well-separated by ion-pair reverse-phase ultra-performance liquid chromatography. Each disaccharide can then be detected and its relative abundance quantified using electrospray ionization mass spectrometry in the negative mode. Because of its high sensitivity, without interference from proteins and other sample impurities, this method is particularly useful in the analysis of low content GlcNH3(+) residues in small amounts of biological materials, eg. sera, tissue and cell culture-derived samples. This would lead to a better understanding of the biological origin of GlcNH3(+) residues and their increasingly important function in human health and disease.
Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, PR China.
Jia Yan
JY
Du
Clinical Laboratory, Fu Zhou First Hospital, Fu Zhou, PR China.
Lan Rong
LR
Chen
Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, PR China.
Su
S
Liu
Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, PR China.
Jiang Hui
JH
Lin
Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, PR China.
Qun Tao
QT
Liang
Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, PR China.
Malcolm
M
Lyon
Institute of Glycobiochemistry, National Engineering Research Centre of Chemical Fertilizer Catalyst, Fu Zhou University, Fu Zhou 350002, PR China. Electronic address: zheng0wei@hotmail.com.
Zheng
Z
Wei
D000818:Animals; D000925:Anticoagulants / Q000737:chemistry*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004187:Disaccharides / Q000032:analysis*; D006493:Heparin / Q000737:chemistry*; D006497:Heparan Sulfate / Q000737:chemistry*; D006801:Humans; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013552:Swine
D000925:Anticoagulants; D004187:Disaccharides; D006493:Heparin; D006497:Heparan Sulfate
Heparan sulfate; Heparin; LC–MS-ITTOF; N-unsubstituted disaccharide