Separation of Vitamins in Mixed-Mode HPLC

Separation of Vitamin B2 and Vitamin C is demonstrated on a Primesep D mixed-mode column. This column can be used for analysis of other hydrophobic or neutral vitamins and supplements.


Column Primesep D , 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer Acetic Acid
Flow Rate 1.0 ml/min
Detection UV, 250 nm


Class of Compounds Drug, Vitamin B₆, Hydrophobic, Ionizable
Analyzing Compounds Vitamins C (ascorbic acid) and B2 (riboflavin)

Application Analytes:

Vitamin B2 (Riboflavin)
Vitamin C