HPLC Analysis of Fat Soluble Vitamins by Normal Phase Chromatography on Ascentis^® Si, 15 cm Column

column

Ascentis Si, 15 cm × 4.6 mm I.D., 5 μm particles (581512-U)

mobile phase

[A] hexane, [B] ethyl acetate

gradient

10 to 30% B in 10 min; held for 2 min; to 10% B in 1 min

flow rate

1.0 mL/min

column temp.

30 °C

detector

UV, 290 nm

injection

10 μL

sample

as indicated in 96:4 (hexane:isopropanol)