Reversed-Phase High-Performance Liquid Chromatography: A Robust Method for Quantitation of Fluorescently Labeled and Derivatized Sialic Acids Isolated From Mouse Liver

Analyze the samples, using a standard HPLC system connected to an online fluorescence detector. Use a reversed-phase C-18 column with a 250-millimeter length and 4.6-millimeter diameter for the analysis.

After preparing solvent A and solvent B, and setting up the HPLC run as detailed in the text protocol, inject 50 microliters of the sample into the HPLC system.

Monitor eluents, using the fluorescence detector excitation wavelength of 373 nanometers, and an emission wavelength of 448 nanometers.

Finally, calculate the relative amount of Neu5Gc as described in the text protocol.