High-performance liquid chromatographic separation of unsaturated disaccharides derived from heparan sulfate and heparin

Lee, George Jia-Long; Tieckelmann, Howard

doi:10.1016/s0021-9673(00)81476-4

Cited by 24 publications

(4 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…RPIP (66) entails addition of millimolar concentrations of an amine to the LC mobile phase to cause pairing of alkyl ammonium cations with the negatively charged GAG oligosaccharides so as to facilitate their interaction with a stationary reversed phase column (67). The amine reagent must be sufficiently volatile to be compatible with ESI MS while providing sufficient hydrophobicity for interaction with the stationary reversed phase.…”

Section: Analysis Of Gags Using Esi Mass Spectrometrymentioning

confidence: 99%

Analysis of Glycosaminoglycans Using Mass Spectrometry

Staples

Zaia

2011

View full text Add to dashboard Cite

The glycosaminoglycans (GAGs) are linear polysaccharides expressed on animal cell surfaces and in extracellular matrices. Their biosynthesis is under complex control and confers a domain structure that is essential to their ability to bind to protein partners. Key to understanding the functions of GAGs are methods to determine accurately and rapidly patterns of sulfation, acetylation and uronic acid epimerization that correlate with protein binding or other biological activities. Mass spectrometry (MS) is particularly suitable for the analysis of GAGs for biomedical purposes. Using modern ionization techniques it is possible to accurately determine molecular weights of GAG oligosaccharides and their distributions within a mixture. Methods for direct interfacing with liquid chromatography have been developed to permit online mass spectrometric analysis of GAGs. New tandem mass spectrometric methods for fine structure determination of GAGs are emerging. This review summarizes MS-based approaches for analysis of GAGs, including tissue extraction and chromatographic methods compatible with LC/MS and tandem MS.

show abstract

Section: Analysis Of Gags Using Esi Mass Spectrometrymentioning

confidence: 99%

Analysis of Glycosaminoglycans Using Mass Spectrometry

Staples

Zaia

2011

View full text Add to dashboard Cite

show abstract

“…The inclusion of quaternary ammonium salts in the mobile phase enables direct separation of GAG disaccharides using a reversed phase chromatography column (Lee & Tieckelmann, 1980). Such reversed phase ion pairing chromatography systems remain popular for disaccharide analysis, particularly when combined with fluorescence-based detection.…”

Section: Lc/ms Of Gagsmentioning

confidence: 99%

On‐line separations combined with MS for analysis of glycosaminoglycans

Zaia

2008

Mass Spectrometry Reviews

116

View full text Add to dashboard Cite

The glycosaminoglycan (GAG) family of polysaccharides includes the unsulfated hyaluronan and the sulfated heparin, heparan sulfate, keratan sulfate, and chondroitin/dermatan sulfate. GAGs are biosynthesized by a series of enzymes, the activities of which are controlled by complex factors. Animal cells alter their responses to different growth conditions by changing the structures of GAGs expressed on their cell surfaces and in extracellular matrices. Because this variation is a means whereby the functions of the limited number of protein gene products in animal genomes is elaborated, the phenotypic and functional assessment of GAG structures expressed spatially and temporally is an important goal in glycomics. On-line mass spectrometric separations are essential for successful determination of expression patterns for the GAG compound classes due to their inherent complexity and heterogeneity. Options include size exclusion, anion exchange, reversed phase, reversed phase ion pairing, hydrophilic interaction, and graphitized carbon chromatographic modes and capillary electrophoresis. This review summarizes the application of these approaches to on-line MS analysis of the GAG classes.

show abstract

“…Heparin lyase, which is commercially available, is optimally active at pH 7.0, the temperature optimum being 42°C. Disaccharides formed by the action of heparin lyase or by the combined action of this enzyme and of heparan sulfate lyase (see the following section) can be quantitated by their absorbance at 232 nm and separated by paper chromatography (469) or high performance liquid chromatography (470).…”

Section: Heparin Lyasementioning

confidence: 99%

“…Calcium ions stabilize the enzyme. As mentioned in the preceding paragraph, the disaccharide products may be separated and quantitated by paper chromatography or high performance liquid chromatography (469,470).…”

Section: Heparan Sulfate Lyasesmentioning

confidence: 99%