Sialic Acid Speciation Using Capillary Electrophoresis: Optimization of Analyte Derivatization and Separation

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• 作者：Zoltan Szabo ; Jonathan Bones ; Andras Guttman ; James Glick ; Barry L. Karger
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：September 18, 2012
• 年：2012
• 卷：84
• 期：18
• 页码：7638-7642
• 全文大小：293K
• 年卷期：v.84,no.18(September 18, 2012)
• ISSN：1520-6882

文摘

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.