Multi-cationic aminopyrene-based tag for oligosaccharide analysis by capillary electrophoresis with
                               laser-induced fluorescence detection



                Denisa Smolková^1,2, Filip Duša^1, Richard Čmelík^1, Jana Lavická^1


^1Institute of Analytical Chemistry of the Czech Academy of Science, Veveří 97, 602 00 Brno, Czech
                                             Republic

      ^2Department of Chemistry, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic


e-mail: smolkova@iach.cz



Glycans and oligosaccharides have a great potential in medical diagnostics, but their analysis is
challenging due to the complexity and variability of the molecules. Oligosaccharides have a low
ionization efficiency important for mass spectrometry and do not contain chromophores or
fluorophores for UV/Vis or fluorescence detection. The way to solve these issues is labeling of
oligosaccharides with fluorescent labels [1] and their analysis by capillary electrophoresis with
laser-induced fluorescence detection (CE/LIF). CE/LIF has the potential as a sensitive and simple
method. The most widely used and commercially available fluorescent label is the trisodium salt of
8-aminopyrene-1,3,6-trisulfonic acid (APTS) [2]. Recently, we have synthetized a new multi-cationic
aminopyrene-based label, APTMP (4,4’,4’’-(8-aminopyrene-1,3,6-trisulfonyl)tris(1-methylpiperazine)
[3]. In this poster, we present optimization of the labeling and separation conditions to achieve
better repeatability and efficiency of labeling as well as detection sensitivity of CE/LIF analysis
of APTMP labeled oligosaccharides. Various reaction conditions were tested for example reaction
temperature, a type of the reducing agent, a reducing agent/fluorescent tag ratio, and an
oligosaccharide/fluorescent tag ratio. The CE/LIF analysis was optimized by testing various
background electrolytes (BGE), capillary coatings, separation voltages, and other experimental
parameters.



Acknowledgements

This work was supported by the Grant Agency of the Czech Republic [22-00236S] and the institutional
research plan [RVO: 68081715].


References

[1] L. Hajba, E. Csanky, A. Guttman, Anal. Chim. Acta 943 (2016) 8–16.

[2] P. Rudd, N.G. Karlsson, K-H. Khoo, N.H. Packer, in: A. Varki, R.D. Cummings, J.D. Esko, (Eds.),
Essentials of Glycobiology, Cold Spring Harbor Laboratory Press, New York, 3rd ed., 2017, 653-666.

[3] J. Krenkova, M. Liskova, R. Cmelik, G. Vigh, F. Foret, Anal. Chim. Acta 1095 (2020)    226–232.