Countercurrent chromatography separation of vitamin E isomers in a co‐current mode

Vitamin E represents a group of lipophilic phenolic compounds, including α‐tocopherol, β‐tocopherol, γ‐tocopherol, and δ‐tocopherol, and α‐tocotrienol, β‐tocotrienol, γ‐tocotrienol, and δ‐tocotrienol isomers. Different forms of vitamin E have been proven to exhibit varying biological activities. However, due to their structural similarities, the separation of vitamin E isomers is a challenging task. Therefore, it is crucial to establish an efficient method for isolating individual isomers. In this study, co‐current countercurrent chromatography was employed to isolate vitamin E isomers from commercial capsules using a n‐heptane‐methanol‐water (10:9.5:0.5, v/v) solvent system. The partition coefficients of the main constituents in the capsules ranged from 0.94 to 6.23, requiring over 450 min for a complete separation. To improve separation efficiency, a co‐current elution mode was implemented and the flow rates of the two liquid phases as well as sample amount were examined. The results suggested that increasing the flow rate of the stationary phase and sample size could result in more effective separation, shorter separation time, and higher yield. It proved that co‐current countercurrent chromatography was an effective method for the separation of vitamin E isomers.