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)
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Application Analytes:
Vitamin B2 (Riboflavin)
Vitamin C

Application Detection:
UV Detection