HPLC Separation and Retention of Vitamins in Mixed-Mode on Zodiac HST B Column

The Zodiac HST B column provides effective separation of Vitamin C (ascorbic acid) and Vitamin B2 (riboflavin) with easily adjustable selectivity. By simply modifying the concentration of acetonitrile in the mobile phase, the elution order of the two vitamins can be reversed. Vitamin B2, in particular, shows high sensitivity to changes in acetonitrile levels. The method employs a mobile phase composed of water, acetonitrile (MeCN), and either trifluoroacetic acid (TFA) or acetic acid (HOAc), with UV detection at 250 nm, delivering excellent resolution and peak performance.

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