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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.
Anal Biochem. 1999 May 01; 269(2):367-78.AB

Abstract

A series of disaccharides derived from chondroitin sulfate and heparin/heparan sulfate were derivatized at their reducing ends with a fluorophore 2-aminobenzamide to develop a sensitive microanalytical method for glycosaminoglycans. The resulting labeled compounds derived from chondroitin sulfate or heparin/heparan sulfate were well-separated and quantified by HPLC equipped with a fluorescence detector. The detection limit was a low picomole level. This method was applied to the analysis of the disaccharide composition of tetra- and hexasaccharides derived from chondroitin sulfate and heparin/heparan sulfate as well as these glycosaminoglycan polysaccharides. The method was also successfully applied to the exosequencing of chondrohexasaccharides, where the fluorophore-labeled oligosaccharides were degraded exolytically from the nonreducing ends using bacterial eliminases. The resultant labeled fragments were identified by HPLC.

Authors+Show Affiliations

Kinoshita A
Department of Biochemistry, Kobe Pharmaceutical University, Kobe, Higashinada-ku, 658-8558, Japan.
Sugahara K
No affiliation info available

MeSH

Carbohydrate SequenceChondroitin SulfatesChromatography, High Pressure LiquidDisaccharidesFluorescent DyesGlycosaminoglycansHeparinHeparitin SulfateMolecular Sequence DataOligosaccharidesSensitivity and Specificityortho-Aminobenzoates

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10222012

Citation

Kinoshita, A, and K Sugahara. "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." Analytical Biochemistry, vol. 269, no. 2, 1999, pp. 367-78.
Kinoshita A, Sugahara K. 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. Anal Biochem. 1999;269(2):367-78.
Kinoshita, A., & Sugahara, K. (1999). 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. Analytical Biochemistry, 269(2), 367-78.
Kinoshita A, Sugahara K. 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. Anal Biochem. 1999 May 1;269(2):367-78. PubMed PMID: 10222012.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Kinoshita,A, AU - Sugahara,K, PY - 1999/5/1/pubmed PY - 1999/5/1/medline PY - 1999/5/1/entrez SP - 367 EP - 78 JF - Analytical biochemistry JO - Anal Biochem VL - 269 IS - 2 N2 - A series of disaccharides derived from chondroitin sulfate and heparin/heparan sulfate were derivatized at their reducing ends with a fluorophore 2-aminobenzamide to develop a sensitive microanalytical method for glycosaminoglycans. The resulting labeled compounds derived from chondroitin sulfate or heparin/heparan sulfate were well-separated and quantified by HPLC equipped with a fluorescence detector. The detection limit was a low picomole level. This method was applied to the analysis of the disaccharide composition of tetra- and hexasaccharides derived from chondroitin sulfate and heparin/heparan sulfate as well as these glycosaminoglycan polysaccharides. The method was also successfully applied to the exosequencing of chondrohexasaccharides, where the fluorophore-labeled oligosaccharides were degraded exolytically from the nonreducing ends using bacterial eliminases. The resultant labeled fragments were identified by HPLC. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/10222012/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_ DB - PRIME DP - Unbound Medicine ER -