HPLC Method For Analysis of Ergocalciferol (Vitamin D2) and Cholecalciferol (Vitamin D3) on Zodiac HST-SB Column

Ergocalciferol (Vitamin D₂) and Cholecalciferol (Vitamin D₃) are fat-soluble vitamins essential for calcium and phosphorus absorption. While the body can synthesize both when exposed to sunlight, they are also commonly obtained through multivitamins or standalone Vitamin D supplements.
Vitamins D₂ and D₃ absorb in the high UV range and can be effectively separated using a Zodiac HST SB reverse-phase column. The method employs a mobile phase of water and acetonitrile, buffered with acetic acid, allowing for efficient retention and resolution. Detection is performed by UV at 265 nm.