High pressure liquid chromatographic separation and identification of vitamins D2 and D3 in the presence of fat-soluble vitamins in dosage forms.
A simple and rapid qualitative method is described for determining the presence of vitamin D2 (ergocalciferol) and/or vitamin D3 (cholecalciferol) in various preparations by reverse phase high pressure liquid chromatography (HPLC). When both D2 and D3 are present, this method effectively separates and identifies each vitamin D form by its respective retention time. A significant difference between vitamins D2 and D3 exists in their antirachitic activity in poultry. Preparations can be tested rapidly by this method to ascertain that the correct D vitamin form has been added. Fat-soluble vitamins such as vitamins A, E, K1, and K3 do not interfere. Vitamins D2 and D3 were separated at the baseline in model preparations. As little as 2 ng each of vitamin D2 and vitamin D3 can be separated and identified.
D002762:Cholecalciferol / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D004304:Dosage Forms; D004872:Ergocalciferols / Q000302:isolation & purification*; D005223:Fats; D007202:Indicators and Reagents; D012995:Solubility
D004304:Dosage Forms; D004872:Ergocalciferols; D005223:Fats; D007202:Indicators and Reagents; D002762:Cholecalciferol