Separation and Retention of Vitamins in Mixed-Mode HPLC

Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Both columns allow peak order reversal by simply changing the acetonitrile concentration in the mobile phase. Vitamin B2 is especially sensitive to acetonitrile concentration. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either trifluoroacetic acid (TFA) or acetic acid (HOAc) with UV detection at 250 nm.