HPLC Analysis of Fat-Soluble Vitamins on Ascentis^® Express C18, Methanol Mobile Phase

column

Ascentis Express C18, 15 cm x 4.6 mm I.D., 2.7 μm particles (53829-U)

mobile phase

methanol

flow rate

1.5 mL/min

pressure

3249 psi (224 bar)

column temp.

30 °C

detector

UV, 280 nm

injection

5 μL

sample

0.5 mg/mL tocopherol acetate, alpha-tocopherol, delta-tocopherol and K1, 0.05 mg/mL D2, D3 and retinol acetate, 1.5 mg/mL retinol palmitate, all in ethanol.