Multimode High-Performance Liquid Chromatography of Fluorescently Labeled Oligosaccharides from Glycoproteins

Townsend, R. Reid; Lipniunas, Peter; Bigge, Collette; Ventom, Andrew M.; Parekh, Raj B.

doi:10.1006/abio.1996.0315

Cited by 44 publications

(27 citation statements)

References 10 publications

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“…Serum (10 L) was denatured in 1% SDS and glycans released with PNGaseF and purified on graphitized carbon (12). A portion was fluorescently labeled at the reducing end with 2-aminobenzamide (2-AB) (13) and separated into neutral and anionic glycans by HPLC (14). Glycan profiles were determined from eight control sera and from a pool of 10 different sera.…”

Section: Methodsmentioning

confidence: 99%

Cryptogenic Liver Disease in Four Children: A Novel Congenital Disorder of Glycosylation

et al. 2006

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ABSTRACT:We investigated the metabolic defect(s) of four children who presented with isolated cryptogenic chronic liver disease, coagulopathy, and abnormalities of several unrelated serum glycoproteins. Analysis of the patients' serum glycoproteins and fibroblasts suggest they have a novel congenital disorder of glycosylation (CDG). All had abnormal transferrin (Tf) isoelectric focusing (IEF) profiles. More detailed analysis of Tf by electrospray ionization mass spectrometry (ESI-MS) showed a plethora of abnormal glycosylations that included loss of 1-2 sialic acids and 1-2 galactose units, typical of Group II defects. Tf from two patients also lacked 1-2 entire oligosaccharide chains, typical of Group One disorders. Total serum N-glycans were analyzed by HPLC and matrix-assisted laser desorption/ionization mass spectrometry and also showed increased proportion of neutral glycan chains lacking sialic acids and galactose units. Analysis of patient fibroblasts eliminated CDG-Ia, through CDG-Ih, -IL and CDG-IId. Our results suggest that a subset of children with clinically asymptomatic, cryptogenic hypertransaminasemia and/or liver steato-fibrosis may represent a novel type of CDG-X with an unknown defect(s). Clinicians are encouraged to test such patients for abnormal Tf glycosylation by ESI-MS.

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Section: Methodsmentioning

confidence: 99%

Cryptogenic Liver Disease in Four Children: A Novel Congenital Disorder of Glycosylation

et al. 2006

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“…An aliquot (5 l) of a derivatization reagent mixture (0.35 M 2AB/1.0 M NaCNBH 4 /30% (v/v) acetic acid in dimethyl sulfoxide) was added to the oligosaccharide sample, and the mixture was incubated at 65°C for 2 h. The derivatized oligosaccharide was purified by paper chromatography using Whatman 3MM paper in a solvent system of butanol:ethanol:water (4:1:1, v/v). Alternatively, the reaction mixture was spotted on a paper strip, which was washed with 1 ml of acetonitrile five times by centrifugation according to the method of Townsend et al (24).…”

Section: Derivatization Of Reducing Oligosaccharides With 2abmentioning

confidence: 99%

Microanalysis of Glycosaminoglycan-Derived Oligosaccharides Labeled with a Fluorophore 2-Aminobenzamide by High-Performance Liquid Chromatography: Application to Disaccharide Composition Analysis and Exosequencing of Oligosaccharides

Kinoshita

Sugahara

1999

Analytical Biochemistry

202

176

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“…Good results for analysis of oligosaccharides was obtained by multimode chromatography (i.e. HILIC) with volatile mobile phases for MALDI/MS [35].…”

Section: Other Polar Stationary Phasesmentioning

confidence: 99%

Application of hydrophilic interaction chromatography in phytochemical analysis

Petruczynik¹,

Waksmundzka‐Hajnos²

2013

Acta Chromatographica

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Summary. Hydrophilic interaction chromatography (HILIC) is a liquid chromatographytechnique when a polar stationary phases -silica or polar bonded phases and aqueous mobile phase containing amount of a less polar solvent (often acetonitrile) is applied. HILIC is important for the separation of highly polar substances including biologically active compounds, such as drugs, amino acids, peptides, proteins, nucleosides, neurotransmitters, etc. HILIC is also the appropriate method for analysis of plant extract polar components such as carbohydrates, amino acids, peptides, phenolic acids, flavonoids and some alkaloids. Plant extracts are usually multicomponent mixtures of compounds of wide polarity range, which often cannot be analyzed by use of a single separation and detection method because of the high chemical diversity. Good results are obtained by use of coupled method, e.g., reversed-phase liquid chromatography (RP-LC) and HILIC mode separation. The elution order in HILIC is usually opposite to that in RP separations. This orthogonality determines one of the advantages of HILIC -the possibility of creating multidimensional separations. An important feature of HILIC is the improved sensitivity with electrospray mass spectrometry. This is significant for the analysis of components existing in small concentration in multicomponent mixtures. The high acetonitrile content also gives additional advantage of HILIC -faster separations due to the lower viscosity of HILIC eluents compared to standard RP ones. The presented review deals with the optimization of HILIC separations and application of the method for analysis of plant extracts components.

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Multimode High-Performance Liquid Chromatography of Fluorescently Labeled Oligosaccharides from Glycoproteins

Cited by 44 publications

References 10 publications

Cryptogenic Liver Disease in Four Children: A Novel Congenital Disorder of Glycosylation

Cryptogenic Liver Disease in Four Children: A Novel Congenital Disorder of Glycosylation

Microanalysis of Glycosaminoglycan-Derived Oligosaccharides Labeled with a Fluorophore 2-Aminobenzamide by High-Performance Liquid Chromatography: Application to Disaccharide Composition Analysis and Exosequencing of Oligosaccharides

Application of hydrophilic interaction chromatography in phytochemical analysis

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