Sialic acid speciation using capillary electrophoresis: Optimization of analyte derivatization and separation

Zoltan Szabo, Jonathan Bones, A. Guttman, James Glick, Barry L. Karger

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Capillary electrophoresis with laser induced fluorescence detection (CE-LIF) was employed for rapid sialic acid speciation, facilitating the quantitative determination of N-glycolylneuraminic acid (Neu5Gc) and N-acetylneuraminic acid (Neu5Ac) on glycoproteins. Derivatization of the sialic acids with 2-aminoacridone (2-AMAC), using classical reductive amination in a nonaqueous solvent, led to the spontaneous decarboxylation of the sialic acid residues as determined by CE-LIF and offline mass spectrometric analysis. Modification of both the labeling conditions, to drive the decarboxylation reaction to completion and the CE-LIF parameters to separate the neutral species by complexation with a neutral coated capillary and borate reversed polarity, led to a robust platform for the rapid, sensitive, and quantitative speciation of sialic acids. The method can readily be used for quality control of recombinant biopharmaceuticals.

Original languageEnglish
Pages (from-to)7638-7642
Number of pages5
JournalAnalytical Chemistry
Volume84
Issue number18
DOIs
Publication statusPublished - Sep 18 2012

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Capillary electrophoresis
N-Acetylneuraminic Acid
Sialic Acids
Fluorescence
Lasers
Amination
Borates
Complexation
Labeling
Quality control
Glycoproteins
Decarboxylation

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Sialic acid speciation using capillary electrophoresis : Optimization of analyte derivatization and separation. / Szabo, Zoltan; Bones, Jonathan; Guttman, A.; Glick, James; Karger, Barry L.

In: Analytical Chemistry, Vol. 84, No. 18, 18.09.2012, p. 7638-7642.

Research output: Contribution to journalArticle

Szabo, Zoltan ; Bones, Jonathan ; Guttman, A. ; Glick, James ; Karger, Barry L. / Sialic acid speciation using capillary electrophoresis : Optimization of analyte derivatization and separation. In: Analytical Chemistry. 2012 ; Vol. 84, No. 18. pp. 7638-7642.
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