Sugar analysis column, Asahipak NH2P-40 2D, was used with a LC/MS to separate N-Acetylchitooligosaccharides. By adding ammonia to the solvent, it enhances the ionization of saccharides, and thus provides high sensitivity detection.
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Sample : 5 μL, Mixture of N-Acetylchitooligosaccharides
(Seikagaku Co. code:800104 Chitooligosaccharides Mixture) 500 ng/mL

1. N-Acetyl-D-glucosamine
2. di-N-Acetylchitobiose
3. tri-N-Acetylchitotriose
4. tetra-N-Acetylchitotetraose
5. penta-N-Acetylchitopentaose
6. hexa-N-Acetylchitohexaose

Column       : Shodex Asahipak NH2P-40 2D (2.0 mm I.D. x 150 mm)
Eluent       : (A); 0.05 % Ammonia aq.
               (B); CH3CN
               Linear gradient; 
               (B %) 75 % (0 min) -> 50 % (10 min) -> 50 % (15 min)
Flow rate    : 0.2 mL/min
Detector     : ESI-MS (SIM Negative)
Column temp. : 30 °C