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Chromatogram

Description

Trisaccharides can be difficult to separate using conventional HPLC columns. The four trisaccharides panose, raffinose, isomaltotriose, and kestose have the same chemical formula, and the all have two glycosidic bonds connecting the three monosaccharides. Obelisc N was used as a stationary phase to separate trisaccharides because it is capable of multiple modes of separation. Obelisc N is a highly polar column that retains polar and charged analytes. Even though the trisaccharides differ only in regio- and stereochemistry they are resolved.

Method Parameters

Mobile PhaseMeCN – 75%
BufferAmFm pH 3.0- 10 mM
Flow Rate1.0 ml/min
DetectionELSD
Class of CompoundsTrisaccharides,  Hydrophilic, Ionizable
Analyzing CompoundsRaffinose, Kestose, Panose, Isomaltose

HPLC Column Used

Obelisc R, 4.6×150 mm, 5 µm, 100A