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Abnormal Behavior of Chromatogram of Saccharides Using a Pb Type Column for Ligand Exchange Chromatography

It is recommended to be analyzed under high temperature (about 80°C) in order to suppress the anomer separation in the analysis of saccharides by ligand exchange mode. When some saccharides are analyzed using Pb type (as counter ion) columns, small bulge might occur near the main peak. This phenomenon is considered to be due to a part of saccharide has been structural change in the column. In this study, fructose was analyzed using SUGAR SP0810. There is a bulge in the rise of the main peak of fructose, but when plotting the peak area including bulge part vs. concentration, highly linear calibration curve is obtained.


Sample : 10µL
Fructose 5000µg/mL

 

Column       : Shodex SUGAR SP0810 (8.0mmI.D. x 300mm)
Eluent       : H2O
Flow rate    : 0.5mL/min
Detector     : Shodex RI
Column temp. : 80°C