HPLC Separation of Vitamin E and Vitamin E Acetate on Primesep B Column
Primesep B separates Vitamin E and Vitamin E acetate with reversed peak order of a traditional C8 column. The different selectivity is due to an embedded basic functional group on the stationary phase which contributes polar interactions to the separation. Excellent peak shape results with a mass spec compatible mobile phase of water and acetonitrile (MeCN, ACN) with UV detection at 205 nm.
Mixed-mode HPLC columns allow for separation of compounds with drastically different properties. In this application, highly hydrophilic ascorbic acid is retained and separated from highly hydrophobic retinol and Vitamin E Acetate. Ascorbic acid is retained by anion-exchange mechanism and hydrophobic compounds are retained by reversed-phase mechanism. Compounds are monitored by UV detection. Mobile phase is compatible with LC/MS and method can be used to monitor basic and acidic compounds in bio-fluids (serum, blood, urine, saliva, etc.)
retinol and Vitamin E Acetate
Condition
Column
Primesep SB , 3.0x50 mm, 5 µm, 100A
Mobile Phase
MeCN/H2O
Buffer
Acetic Acid
Flow Rate
0.5 ml/min
Detection
UV 270, 325
Description
Class of Compounds
Acid, Vitamin B₆, Hydrophobic, Ionizable
Analyzing Compounds
Ascorbic acid (Vitamin C), Retinol and Vitamin E Acetate