Separation of vitamins retinol acetate, ergocalciferol, or choloecalciferol and tocopherol acetate using sequential injection chromatography
2011
Abstract
A novel simple method for separation of vitamins A-acetate (all-trans retinol acetate), D2 (ergocalciferol), or D3 (cholecalciferol) and E-acetate (tocopherol acetate) using short monolithic column in the sequential injection chromatography system is described. Separation was carried out using FIAlab (R) 3000 system under following conditions: Onyx Monolithic C18 column (25x4.6mm), mobile phase acetonitrile:methanol:H2O 20:20:1 (v/v/v)), flow rate 0.9mLmin-1, detection at 265nm (D), 290nm (E), and 325nm (A).
The method was validated with respect to peak asymmetry, resolution, number of theoretical plates, repeatability, linearity, precision, and accuracy. Analysis time was 5.5 min.