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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 6E ( 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(0.1μg)/mL each, 100μL
1. Vitamin D2(Ergocalciferol)
2. Vitamin D3(Cholecaciferol)



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