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Electrophoretic Separations of Oligosaccharides Coupled with On-line Fluorescent Labelling

Autoři: Nováková Zuzana | Česla Petr | Fischer Jan

Rok: 2016

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	Electrophoretic Separations of Oligosaccharides Coupled with On-line Fluorescent Labelling	Carbohydrates, either alone or as constituents of glycoproteins and glycolipids, are mediators of several events, such as intra and extracellular recognition, differentiation, proliferation or even signal transduction. Oligosaccharides play a significant role in posttranslational modifications of proteins. The changes in protein glycosylation may serve as an indicator of some diseases. Carbohydrate derivatization is required for most of the analytical procedures. The determination of glycans (without treatment) presented in organisms is very difficult. Detection of intact carbohydrates can be utilized by spectroscopic measurement at 195 nm, amperometric or refractive index detection, all of which are associated with low sensitivities. Derivatization with UV-absorbing or fluorescent molecules significantly enhances the detection sensitivity of highperformance liquid chromatography (HPLC), capillary electrophoresis (CE) and polyacrylamide gel electrophoresis (PAGE). It is necessary to insert an optically active substance into the analyzed molecule. For example, maltooligosaccharides maltose to maltoheptaose) can be labelled with 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS) and used for their characterization. In present work, conditions in non-coated and coated capillaries were optimized. Further, we had examined and compared various fluorescent derivatization agents in terms of resolution, migration time, and sensitivity of detection. Finally, we have developed the method for on-line labelling of oligosaccharides using procedure for fluorescent derivatization.

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