Polymer-based amino column, Asahipak NH2P-50 4E, was used to analyze water-soluble vitamins under HILIC mode. Six water-soluble vitamins were separated within 5 minutes.

[Note] This analysis requires column equilibration.
For column equilibration: First introduce 60 mM phosphoric acid at 0.5 mL/min for 2 hours, and then switch to the eluent.


Sample : 20 μL
1. Vitamin B₆ 50 μg/mL
2. Nicotinamide 10 μg/mL
3. Vitamin B₁₂ 10 μg/mL
4. Nicotinic acid 10 μg/mL
5. Folic acid 10 μg/mL
6. Vitamin C 10 μg/mL

Column       : Shodex Asahipak NH2P-50 4E (4.6 mm I.D. x 250 mm)
Eluent       : 40 mM H3PO4 aq./CH3CN=45/55 
Flow rate    : 1.0 mL/min
Detector     : UV (254 nm)
Column temp. : 40 °C