HPLC Separation of Vitamin C and Vitamin B2

Vitamin C (ascorbic acid) and Vitamin B2 (riboflavin) are essential water-soluble vitamins that are not stored in the body and therefore must be replenished daily. The Zodiac HST RP mixed-mode column provides efficient resolution of both vitamins.

The elution order of Vitamin C and Vitamin B2 can be reversed by adjusting the acetonitrile and acetic acid concentrations in the mobile phase, offering enhanced method flexibility.

The HPLC separation is performed using a mobile phase of water, acetonitrile (MeCN/ACN), and acetic acid (AcOH) with UV detection at 250 nm, enabling robust analysis of these vitamins in nutritional and pharmaceutical formulations.