Vitamin D₂ and D₃ were separated using Asahipak ODP-50 6E, a polymer-based reversed phase column. ODP-50 has a wide pH range and can be washed with an aqueous solution of acid or alkali, so it is a highly durable column even for samples with a lot of impurities.


Sample : 4 IU (0.1 μg)/mL each, 100 μL
1. Vitamin D₂ (Ergocalciferol)
2. Vitamin D₃ (Cholecaciferol)

Column       : Shodex Asahipak ODP-50 6E (6.0 mm I.D. x 250 mm) 
Eluent       : CH3CN/CH3OH=90/10                       
Flow rate    : 1.5 mL/min
Detector     : UV (265 nm)
Column temp. : 27 °C