Analysis Of Sialic Acid By High-performance Liquid Chromatography With Fluorescence Detection

Sialic acids are a monosaccharide family，which are widespread in animals. Sialicacids are frequently found as the terminal sugars of glycolipids and glycoproteins. Theirlocations lead them to playing an important role in many biochemical reactions in vivo.There are more than50kinds of sialic acids that have been discovered, and somephenomena of physiological and pathological that sialic acids involve have been reported.However, the specific roles of sialic acids in these phenomena are still unknown.Functions of sialic acids are affected by their structures and concentrations. Theapplications for the accurate and quick detection of sialic acids will be significant inbiology. The methods of sialic acids detection commonly include spectroscopy, gaschromatograph, liquid chromatography, mass spectrometry and amperometric method, etc.In recent years, fluorescence detection by high performance liquid chromatography hasgot more and more attentions because of fast separation, high sensitivity and low sampleconsumption.The main work of this paper was that based on the well performance of HPLC onseparation and high-sensitivity of fluorescence detection, a method used for sialic acidsanalysis had been established. In this method,1,2-diamino-4,5-methylenedioxybenzene(DMB) was taken as the pre-column derivative reagent, which could label the sialic acids.Separation was taken by high performance liquid chromatography, and detection wastaken by fluorescence detector with the Ex373nm, Em448nm.The most optimizedconditions for derivatization were obtained by adjusting the reaction temperature, time, theconcentration of fluorescent reagent and analysing the results. Then, regression equation,linear range and limit of detection were calculated. The system was applied in detectingthe concentrations of sialic acids in rat serum and mouse serum.