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Prediction of Gradient Retention Data of Oligomeric Series in HILIC

Authors: Vaňková Nikola | Česla Petr

Year: 2016

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Prediction of Gradient Retention Data of Oligomeric Series in HILIC	In this study, we have investigated mixed-mode retention model and its extended version for predictions of retention times of native and fluorescently labelled oligosaccharides separated in HILIC in gradient elution mode. Native oligosaccharides were labeled using four reagents: 2-aminobenzoic acid, 2 aminobenzamide, 2-aminopyridine and 8-aminonaphthalene-1,3,6-trisulfonic acid. The separations were realized on the column packed with amide-type stationary phase in mobile phases consisted of acetonitrile and water with 0.1% acetic acid (v/v) and acetonitrile with 30 mM ammonium acetate. For description and prediction of retention behavior in HILIC, the mixed-mode isocratic retention model can be used, which can be extended with parameter describing contribution of glucose unit to the retention of native and derivatized maltooligosaccharides. These models are describing retention in isocratic elution. In gradient elution, the mobile phase composition is changing over the time of analysis. For description of retention in gradient elution, both models were used for calculation of immediate value of retention factor of solute band at each volume element dV and numerically integrated. Model parameters used for calculations of theoretical retention times were obtained using both, isocratic and gradient experiments. Retention times of native and derivatized maltooligosaccharadies were measured at 15 different gradient profiles (φ0 (ACN) = 0.05, 0.15, 0.25 φend (ACN) = 0.5, 0.7, 0.8) and tG = 5, 7.5, 10 min. In general the mixed-mode model provided better results compared to the model considering retention of all oligomers, however, only one scouting gradient run is enough for calculation of model parameters in this case. The highest average deviation obtained for mobile phase with 30 mM ammonium acetate calculated for mixed-mode retention model was 10.22 % and for extended model it was 14 %.