Analysis of Fluorescently Labeled Sugars by Reversed-Phase Ion-Pairing High Performance Liquid Chromatography
Authors
Kim, Y.S.
Liu, J.
Han, X.J.
Pervin, A.
Linhardt, Robert J.
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
1995-01-01
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Degree
Terms of Use
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Full Citation
Analysis of Fluorescently Labeled Sugars by Reversed-Phase Ion-Pairing High Performance Liquid Chromatography, Y. S. Kim, J. Liu, X.J. Han, A. Pervin, R. J. Linhardt, Journal of Chromatographic Science, 33, 162-167 1995.
Abstract
Reducing sugars, including monosaccharides, disaccharides, and a trisaccharide, are derivatized by reductive amination with 7-amino-1,3-naphthalene disulfonic acid. Reversed-phase ion-pairing high-performance liquid chromatography is then used to separate these visibly fluorescent, charged conjugates. Isocratic elution with triethylamine-acetic acid from a phenyl column, a C18 column, and C18 and phenyl columns in series gives good separations of a mixture of monosaccharides and a mixture of disaccharides and trisaccharides. Resolution of certain monosaccharides is enhanced by replacing triethylamine with a chiral amine and using gradient elution. Further enhancement of resolution is achieved by adding phenylboronic acid, an agent capable of complexing with the vicinal diol functionality present in many sugars. The trimethylamine-acetic acid eluant permits detection by either ultraviolet absorbance or fluorescence, and the addition of a chiral ion-pairing agent or a phenylboronic acid complexing agent necessitates fluorescence detection. A reversible Schiff base form of the fluorescent sugar conjugate is prepared; it is sufficiently stable to perform fractionations but sufficiently unstable to be converted to a fluorescent label and reducing sugar.
Description
Journal of Chromatographic Science, 33, 162-167
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
Publisher
Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Journal of chromatographic science
https://harc.rpi.edu/
Rensselaer Polytechnic Institute, Troy, NY
Journal of chromatographic science
https://harc.rpi.edu/
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https://login.libproxy.rpi.edu/login?url=https://doi.org/10.1093/chromsci/33.4.162