Direct detection method of oligosaccharides by high-performance liquid chromatography with charged aerosol detection |
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Authors: | Inagaki Shinsuke Min Jun Zhe Toyo'oka Toshimasa |
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Institution: | Division of Bio-Analytical Chemistry, School of Pharmaceutical Sciences, COE Program in the 21st Century, University of Shizuoka, 52-1 Yada, Surugaku, Shizuoka 422-8526, Japan. |
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Abstract: | A simple and rapid detection method of oligosaccharides using high-performance liquid chromatography with a charged aerosol detection (HPLC-CAD) was studied. The direct detection of a sialylglycopeptide (SGP) derived from egg yolk was accomplished by HPLC-CAD using an amido-silica column, and its limit of detection was 0.40 pmol signal-to-noise ratio (S/N) = 3]. The sensitivity of this method was lower than that of the fluorescence detection; however, the method showed approximately 5 times higher sensitivity than that using the conventional UV absorbance detection. Furthermore, this method was used for the analysis of the acid hydrolysis products of SGP. Monosialo- and asialo-oligosaccharides as well as free sialic acid were detected without using fluorescent derivatization. These results indicate that the present method is a new tool for the analysis of oligosaccharides. |
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Keywords: | HPLC hydrophilic interaction liquid chromatography (HILIC) charged aerosol detection (CAD) fluorescent derivatization oligosaccharides |
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