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Fluorescence Detection Speeds Determination of Sialic Acids
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Thermo Fisher Scientific has announced a new ultra high-performance liquid chromatography (UHPLC) method that allows high resolution of sialic acids in a 10 min analysis time and requires less organic solvent than other published methods.
Application Note 278: UHPLC Determination of Sialic Acids with Fluorescence Detection demonstrates that the sensitivity of fluorescence detection easily allows straightforward determination of sialic acids in the pmol range.
In AN 278, N-acetylated sialic acids are determined and O-acetylated sialic acids are identified by UHPLC with fluorescence detection following acid hydrolysis and 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride (DMB) derivatization of glycoprotein samples.
The described assay uses a short format Thermo Scientific Acclaim Rapid Separation LC (RSLC) 120 C18 column that allows fast run times (10 min) and requires less organic solvent compared to a commonly used 40 min method.
The sensitivity of fluorescence detection easily allows determination of sialic acids that are present in the pmol range in glycoprotein hydrolyzates.
This sensitivity allows for straightforward determination of Neu5Ac, Neu5Gc, and O-acetylated sialic acids in the derivatized samples.
This application note and many others can be found at www.thermoscientific.com/dionex under the Documents tab.
