Application Note: Vitamin D2 and D3 Separation by New Highly Hydrophobic UHPLC/HPLC Phase
14 December 2015

Separations of structurally similar compounds such as vitamin D2 and vitamin D3 are very challenging. Vitamin D2 (ergocalciferol) and D3 (cholecalciferol) can be found in different foods including fatty fishes, meat, eggs and some mushrooms. Both compounds are (indirectly) involved in a number of biological functions in the body, including bone metabolism and enhancement of intestinal absorption of calcium, iron, magnesium, phosphate and zinc. A regular intake of vitamin D therefore is essential. Standard C18 columns are not able to fully separate the two vitamins. A very hydrophobic phase with a higher carbon coverage and therefore a greater density of C18 chains is required. This application note demonstrates the use of the YMC-Triart C18 ExRS to separate these compounds.