1 research outputs found
Sialic Acid Speciation Using Capillary Electrophoresis: Optimization of Analyte Derivatization and Separation
Capillary electrophoresis with laser induced fluorescence
detection
(CE-LIF) was employed for rapid sialic acid speciation, facilitating
the quantitative determination of <i>N</i>-glycolylneuraminic
acid (Neu5Gc) and <i>N</i>-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