Vitamin D

Vitamin D's are ususally separated by normal phase mode or reversed phase mode. Vitamin D₂ and D₃ were separated using Asahipak ODP-50 6D ( polymer-based column ) by reversed phase mode, though they could not be separated using Silica 5SIL 4E ( silica-based column ) by normal phase mode. Since the usable pH range of the ODP-50 column is very wide and it can be washed with both acidic and alkaline solvents, it is very durable even if it is used for crude samples.



Sample : 4IU(0.1micro-g)/mL each, 100micro-L
1. Vitamin D₂(Ergocalciferol)
2. Vitamin D₃(Cholecaciferol)

Columns      : Shodex Asahipak ODP-50 6D (6.0mmID*150mm)  Shodex Silica 5SIL 4E (4.6mmID*250mm)
Eluent       : CH3CN/CH3OH=90/10                          Hexane/Isopropanol=99.6/0.4
Flow rate    : 1.5mL/min                                  1.6mL/min
Detector     : UV(265nm)
Column temp. : 27deg-C