HPLC Analysis of Ascorbic Acid, Retinol, and Vitamin E Acetate in ACE Serum Actives Using Zodiac HST-SB Column

Mixed-mode HPLC columns enable the separation of compounds with widely differing physicochemical properties. In this application, the highly hydrophilic ascorbic acid is effectively separated from the highly hydrophobic compounds retinol (Vitamin A) and Vitamin E acetate using a ZodiaA HST SB column. Ascorbic acid is retained primarily through anion-exchange interactions, while the hydrophobic vitamins are retained via reversed-phase mechanisms. All compounds are monitored using UV detection. The mobile phase is fully compatible with LC/MS, making this method suitable for analyzing both acidic and basic compounds in various biological fluids such as serum, blood, urine, and saliva.