文摘
Among glycosaminoglycans, polysulfated heparin chainsprovide the greatest challenge to characterization due tohigh polarity, structural diversity, and sulfate lability.Thepresent report demonstrates how electrospray mass spectrometry (ESMS) can be used to derive compositionalinformation from pure and mixed fractions of heparintetra- to decasaccharide fragments prepared by controlleddigestion of heparin with heparinase I. It alsodescribesan improved procedure for fractionation of heparin oligosaccharides up to octadecasaccharides. Ammoniumsalts prove to be superior to sodium salts, particularlyforanalysis of mixed components. In the mass spectrum ofa hexasaccharide fraction, the identification of six majormass peaks that represent the known hexasaccharidestructures confirms that ESMS analysis of heparin oligosaccharide fragments gives a close representation oftheir constituent composition. In addition to thepreviously identified components, several unreported oligosaccharides were detected in the spectra of octa- anddecasaccharide fractions. The ESMS identification of thethreemajor species in a decasaccharide fraction was confirmedafter HPLC subfractionation and reanalysis. ESMSdetection sensitivity of low picomole amounts of oligosaccharides can be readily achieved.